CaMKII INHIBITORS AND USES THEREOF

ABSTRACT

The present invention provides compounds useful as inhibitors of Ca 2+ /calmodulin-dependent protein kinase (CaMKII), compositions thereof, and methods of using the same.

BACKGROUND OF THE INVENTION

Cardiovascular disease remains the number one cause of death indeveloped countries. Furthermore, incidence of cardiovascular diseasehas increased dramatically in developing countries. Althoughcardiovascular disease usually affects older adults, the antecedents ofcardiovascular disease, notably atherosclerosis, begin in early life,making primary prevention efforts necessary from childhood. Therefore,there is increased emphasis on preventing atherosclerosis by modifyingrisk factors, such as healthy eating, exercise, and avoidance ofsmoking. It is estimated that 1 in 3 people will die from complicationsattributable to cardiovascular disease. “Global Atlas on CardiovascularDisease Prevention and Control”, World Health Organization; January2012. In order to stem the tide and address the shifting epidemiology ofthis disease, measures to prevent or reverse cardiovascular disease mustbe taken.

Obesity and diabetes mellitus are often linked to cardiovasculardisease, as are a history of chronic kidney disease andhypercholesterolemia. In fact, cardiovascular disease is the most lifethreatening of the diabetic complications and diabetics are two- tofour-fold more likely to die of cardiovascular-related causes thannondiabetics.

Diet and exercise, even when used in conjunction with the currentpharmacotherapy, often do not provide sufficient control ofcardiovascular symptoms. The continuing and highly prevalent problem ofcardiovascular disease highlights the overwhelming need for new drugs totreat this condition and its underlying causes.

SUMMARY OF THE INVENTION

It has now been found that compounds of this invention, andpharmaceutically acceptable compositions thereof, are effective asinhibitors of Ca²⁺/calmodulin-dependent protein kinase II (CaMKII). Suchcompounds have the general formula I:

or a pharmaceutically acceptable salt thereof, wherein each variable isas defined and described herein.

Compounds of the present invention, and pharmaceutically acceptablecompositions thereof, are useful for treating a variety of diseases,disorders or conditions. For example, provided compounds are useful intreatment of diseases, disorders or conditions associated with theregulation and inhibition of CaMKII. Such diseases, disorders, orconditions include those described herein.

Compounds provided by this invention are also useful for the study ofCaMKII enzymes in biological and pathological phenomena; the study ofintracellular signal transduction pathways occurring in cardiac,vascular and other bodily tissues; and the comparative evaluation of newCaMKII inhibitors or other regulators of inflammation in vitro or invivo.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 presents results of an in vitro CaMKII enzyme inhibition assayfor compound I-7.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS 1. GENERAL DESCRIPTION OFCOMPOUNDS OF THE INVENTION

In certain embodiments, the present invention provides inhibitors ofCaMKII. In some embodiments, such compounds include those of formula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   one of R^(2′) and R^(3′) is -L¹-R^(x), and the other is hydrogen;-   L¹ is a covalent bond or a straight or branched C₁₋₆ aliphatic    group, wherein one or more methylene groups are independently and    optionally replaced by —NR^(a)— or —O—;-   R^(x) is selected from the group consisting of NH₂, guanidino, 4-7    membered optionally substituted saturated heterocyclic ring having    1-2 heteroatoms independently selected from nitrogen, oxygen or    sulfur, and 5-6 membered heteroaromatic ring having 1-2 heteroatoms    independently selected from sulfur, nitrogen and oxygen;-   each of R³, R⁴, R⁵, R⁶, R⁷, R⁸, R^(4′), R^(5′), and R^(6′) is    independently selected from the group consisting of hydrogen,    halogen, —CN, —CF₃, —OR, —NR₂, —NO₂, —COOR, —CONR₂, and —R;-   each R^(a) is independently hydrogen or C₁₋₃ aliphatic; and-   each R is independently hydrogen or optionally substituted C₁₋₆    aliphatic.

2. COMPOUNDS AND DEFINITIONS

Compounds of this invention include those described generally above, andare further illustrated by the classes, subclasses, and speciesdisclosed herein. As used herein, the following definitions shall applyunless otherwise indicated. For purposes of this invention, the chemicalelements are identified in accordance with the Periodic Table of theElements, CAS version, Handbook of Chemistry and Physics, 75^(th) Ed.Additionally, general principles of organic chemistry are described in“Organic Chemistry”, Thomas Sorrell, University Science Books,Sausalito: 1999, and “March's Advanced Organic Chemistry”, 5^(th) Ed.,Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York: 2001, theentire contents of which are hereby incorporated by reference.

The term “aliphatic” or “aliphatic group”, as used herein, means astraight-chain (i.e., unbranched) or branched, substituted orunsubstituted hydrocarbon chain that is completely saturated or thatcontains one or more units of unsaturation, or a monocyclic hydrocarbonor bicyclic hydrocarbon that is completely saturated or that containsone or more units of unsaturation, but which is not aromatic (alsoreferred to herein as “carbocycle,” “cycloaliphatic” or “cycloalkyl”),that has a single point of attachment to the rest of the molecule.Unless otherwise specified, aliphatic groups contain 1-6 aliphaticcarbon atoms. In some embodiments, aliphatic groups contain 1-5aliphatic carbon atoms. In other embodiments, aliphatic groups contain1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groupscontain 1-3 aliphatic carbon atoms, and in yet other embodiments,aliphatic groups contain 1-2 aliphatic carbon atoms. In someembodiments, “cycloaliphatic” (or “carbocycle” or “cycloalkyl”) refersto a monocyclic C₃-C₆ hydrocarbon that is completely saturated or thatcontains one or more units of unsaturation, but which is not aromatic,that has a single point of attachment to the rest of the molecule.Suitable aliphatic groups include, but are not limited to, linear orbranched, substituted or unsubstituted alkyl, alkenyl, alkynyl groupsand hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or(cycloalkyl)alkenyl.

The term “lower alkyl” refers to a C₁₋₄ straight or branched alkylgroup. Exemplary lower alkyl groups are methyl, ethyl, propyl,isopropyl, butyl, isobutyl, and tert-butyl.

The term “lower haloalkyl” refers to a C₁₋₄ straight or branched alkylgroup that is substituted with one or more halogen atoms.

The term “heteroatom” means one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon (including, any oxidized form of nitrogen,sulfur, phosphorus, or silicon; the quaternized form of any basicnitrogen or; a substitutable nitrogen of a heterocyclic ring, forexample N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) orNR⁺ (as in N-substituted pyrrolidinyl)).

The term “unsaturated,” as used herein, means that a moiety has one ormore units of unsaturation.

As used herein, the term “bivalent C₁₋₈ (or C₁₋₆) saturated orunsaturated, straight or branched, hydrocarbon chain”, refers tobivalent alkylene, alkenylene, and alkynylene chains that are straightor branched as defined herein.

The term “alkylene” refers to a bivalent alkyl group. An “alkylenechain” is a polymethylene group, i.e., —(CH₂)_(n)—, wherein n is apositive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from1 to 2, or from 2 to 3. A substituted alkylene chain is a polymethylenegroup in which one or more methylene hydrogen atoms are replaced with asubstituent. Suitable substituents include those described below for asubstituted aliphatic group.

The term “alkenylene” refers to a bivalent alkenyl group. A substitutedalkenylene chain is a polymethylene group containing at least one doublebond in which one or more hydrogen atoms are replaced with asubstituent. Suitable substituents include those described below for asubstituted aliphatic group.

As used herein, the term “cyclopropylenyl” refers to a bivalentcyclopropyl group of the following structure:

As used herein, the term “cyclobutylenyl” refers to a bivalentcyclobutyl group of the following structure:

As used herein, the term “oxetanyl” refers to a bivalent oxetanyl groupof the following structure:

The term “halogen” means F, Cl, Br, or I.

The term “aryl” used alone or as part of a larger moiety as in“aralkyl,” “aralkoxy,” or “aryloxyalkyl,” refers to monocyclic andbicyclic ring systems having a total of five to 10 ring members, whereinat least one ring in the system is aromatic and wherein each ring in thesystem contains three to seven ring members. The term “aryl” may be usedinterchangeably with the term “aryl ring”. In certain embodiments of thepresent invention, “aryl” refers to an aromatic ring system whichincludes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl andthe like, which may bear one or more substituents. Also included withinthe scope of the term “aryl,” as it is used herein, is a group in whichan aromatic ring is fused to one or more non-aromatic rings, such asindanyl, phthalimidyl, naphthimidyl, phenanthridinyl, ortetrahydronaphthyl, and the like.

The terms “heteroaryl” and “heteroar-,” used alone or as part of alarger moiety, e.g., “heteroaralkyl,” or “heteroaralkoxy,” refer togroups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms;having 6, 10, or 14 π electrons shared in a cyclic array; and having, inaddition to carbon atoms, from one to five heteroatoms. The term“heteroatom” refers to nitrogen, oxygen, or sulfur, and includes anyoxidized form of nitrogen or sulfur, and any quaternized form of a basicnitrogen. Heteroaryl groups include, without limitation, thienyl,furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl,oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl,thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl,purinyl, naphthyridinyl, and pteridinyl. The terms “heteroaryl” and“heteroar-”, as used herein, also include groups in which aheteroaromatic ring is fused to one or more aryl, cycloaliphatic, orheterocyclyl rings, where the radical or point of attachment is on theheteroaromatic ring. Nonlimiting examples include indolyl, isoindolyl,benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl,benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl,quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl,phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Aheteroaryl group may be mono- or bicyclic. The term “heteroaryl” may beused interchangeably with the terms “heteroaryl ring,” “heteroarylgroup,” or “heteroaromatic,” any of which terms include rings that areoptionally substituted. The term “heteroaralkyl” refers to an alkylgroup substituted by a heteroaryl, wherein the alkyl and heteroarylportions independently are optionally substituted.

As used herein, the terms “heterocycle,” “heterocyclyl,” “heterocyclicradical,” and “heterocyclic ring” are used interchangeably and refer toa stable 5- to 7-membered monocyclic or 7-10-membered bicyclicheterocyclic moiety that is either saturated or partially unsaturated,and having, in addition to carbon atoms, one or more, preferably one tofour, heteroatoms, as defined above. When used in reference to a ringatom of a heterocycle, the term “nitrogen” includes a substitutednitrogen. As an example, in a saturated or partially unsaturated ringhaving 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, thenitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as inpyrrolidinyl), or ⁺NR (as in N-substituted pyrrolidinyl).

A heterocyclic ring can be attached to its pendant group at anyheteroatom or carbon atom that results in a stable structure and any ofthe ring atoms can be optionally substituted. Examples of such saturatedor partially unsaturated heterocyclic radicals include, withoutlimitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl,piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl,diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. Theterms “heterocycle,” “heterocyclyl,” “heterocyclyl ring,” “heterocyclicgroup,” “heterocyclic moiety,” and “heterocyclic radical,” are usedinterchangeably herein, and also include groups in which a heterocyclylring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings,such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, ortetrahydroquinolinyl, where the radical or point of attachment is on theheterocyclyl ring. A heterocyclyl group may be mono- or bicyclic. Theterm “heterocyclylalkyl” refers to an alkyl group substituted by aheterocyclyl, wherein the alkyl and heterocyclyl portions independentlyare optionally substituted.

As used herein, the term “partially unsaturated” refers to a ring moietythat includes at least one double or triple bond. The term “partiallyunsaturated” is intended to encompass rings having multiple sites ofunsaturation, but is not intended to include aryl or heteroarylmoieties, as herein defined.

As described herein, compounds of the invention may contain “optionallysubstituted” moieties. In general, the term “substituted,” whetherpreceded by the term “optionally” or not, means that one or morehydrogens of the designated moiety are replaced with a suitablesubstituent. Unless otherwise indicated, an “optionally substituted”group may have a suitable substituent at each substitutable position ofthe group, and when more than one position in any given structure may besubstituted with more than one substituent selected from a specifiedgroup, the substituent may be either the same or different at everyposition. Combinations of substituents envisioned by this invention arepreferably those that result in the formation of stable or chemicallyfeasible compounds. The term “stable,” as used herein, refers tocompounds that are not substantially altered when subjected toconditions to allow for their production, detection, and, in certainembodiments, their recovery, purification, and use for one or more ofthe purposes disclosed herein.

Suitable monovalent substituents on a substitutable carbon atom of an“optionally substituted” group are independently halogen;—(CH₂)₀₋₄R^(o); —(CH₂)₀₋₄OR^(o); —O(CH₂)₀₋₄R^(o), —O—(CH₂)₀₋₄C(O)OR^(o);—(CH₂)₀₋₄CH(OR^(o))₂; —(CH₂)₀₋₄SR^(o); —(CH₂)₀₋₄Ph, which may besubstituted with R^(o); —(CH₂)₀₋₄O(CH₂)₀₋₁Ph which may be substitutedwith R^(o); —CH═CHPh, which may be substituted with R^(o);—(CH₂)₀₋₄O(CH₂)₀₋₁-pyridyl which may be substituted with R^(o); —NO₂;—CN; —N₃; —(CH₂)₀₋₄N(R^(o))₂; —(CH₂)₀₋₄N(R^(o))C(O)R^(o);—N(R^(o))C(S)R^(o); —(CH₂)₀₋₄N(R^(o))C(O)NR^(o) ₂; —N(R^(o))C(S)NR^(o)₂; —(CH₂)₀₋₄N(R^(o))C(O)OR^(o); —N(R^(o))N(R^(o))C(O)R^(o);—N(R^(o))N(R^(o))C(O)NR^(o) ₂; —N(R^(o))N(R^(o))C(O)OR^(o);—(CH₂)₀₋₄C(O)R^(o); —C(S)R^(o); —(CH₂)₀₋₄C(O)OR^(o);—(CH₂)₀₋₄C(O)SR^(o); —(CH₂)₀₋₄C(O)OSiR^(o) ₃; —(CH₂)₀₋₄OC(O)R^(o);—OC(O)(CH₂)₀₋₄SR—, SC(S)SR^(o); —(CH₂)₀₋₄SC(O)R^(o); —(CH₂)₀₋₄C(O)NR^(o)₂; —C(S)NR^(o) ₂; —C(S)SR^(o); —SC(S)SR^(o), —(CH₂)₀₋₄OC(O)NR^(o) ₂;—C(O)N(OR^(o))R^(o); —C(O)C(O)R^(o); —C(O)CH₂C(O)R^(o);—C(NOR^(o))R^(o); —(CH₂)₀₋₄SSR^(o); —(CH₂)₀₋₄S(O)₂R^(o);—(CH₂)₀₋₄S(O)₂OR^(o); —(CH₂)₀₋₄OS(O)₂R^(o); —S(O)₂NR^(o) ₂;—(CH₂)₀₋₄S(O)R^(o); —N(R^(o))S(O)₂NR^(o) ₂; —N(R^(o))S(O)₂R^(o);—N(OR^(o))R^(o); —C(NH)NR^(o) ₂; —P(O)₂R^(o); —P(O)R^(o) ₂; —OP(O)R^(o)₂; —OP(O)(OR^(o))₂; SiR^(o) ₃; —(C₁₋₄ straight or branchedalkylene)O—N(R^(o))₂; or —(C₁₋₄ straight or branchedalkylene)C(O)O—N(R^(o))₂, wherein each R^(o) may be substituted asdefined below and is independently hydrogen, C₁₋₆ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, —CH₂-(5-6 membered heteroaryl ring), or a 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(o), taken together with their intervening atom(s), form a3-12-membered saturated, partially unsaturated, or aryl mono- orbicyclic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, which may be substituted as defined below.

Suitable monovalent substituents on R^(o) (or the ring formed by takingtwo independent occurrences of R^(o) together with their interveningatoms), are independently halogen, —(CH₂)₀₋₂R^(•), -(haloR^(•)),—(CH₂)₀₋₂OH, —(CH₂)₀₋₂OR^(•), —(CH₂)₀₋₂CH(OR^(•))₂; —O(haloR^(•)), —CN,—N₃, —(CH₂)₀₋₂C(O)R^(•), —(CH₂)₀₋₂C(O)OH, —(CH₂)₀₋₂C(O)OR^(•),—(CH₂)₀₋₂SR^(•), —(CH₂)₀₋₂SH, —(CH₂)₀₋₂NH₂, —(CH₂)₀₋₂NHR^(•),—(CH₂)₀₋₂NR^(•) ₂, —NO₂, —SiR^(•) ₃, —OSiR^(•) ₃, —C(O)SR^(•), —(C₁₋₄straight or branched alkylene)C(O)OR^(•), or —SSR^(•) wherein each R^(•)is unsubstituted or where preceded by “halo” is substituted only withone or more halogens, and is independently selected from C₁₋₄ aliphatic,—CH₂Ph, —O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. Suitable divalent substituents on asaturated carbon atom of R^(o) include ═O and ═S.

Suitable divalent substituents on a saturated carbon atom of an“optionally substituted” group include the following: ═O, ═S, ═NNR*₂,═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O)₂R*, ═NR*, ═NOR*, —O(C(R*₂))₂₋₃O—, or—S(C(R*₂))₂₋₃S—, wherein each independent occurrence of R* is selectedfrom hydrogen, C₁₋₆ aliphatic which may be substituted as defined below,or an unsubstituted 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur. Suitable divalent substituents that are bound tovicinal substitutable carbons of an “optionally substituted” groupinclude: —O(CR*₂)₂₋₃O—, wherein each independent occurrence of R* isselected from hydrogen, C₁₋₆ aliphatic which may be substituted asdefined below, or an unsubstituted 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R* include halogen,—R^(•), -(haloR^(•)), —OH, —OR^(•), —O(haloR^(•)), —CN, —C(O)OH,—C(O)OR^(•), —NH₂, —NHR^(•), —NR^(•) ₂, or —NO₂, wherein each R^(•) isunsubstituted or where preceded by “halo” is substituted only with oneor more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

Suitable substituents on a substitutable nitrogen of an “optionallysubstituted” group include —R^(†), —NR^(†) ₂, —C(O)R^(†), —C(O)OR^(†),—C(O)C(O)R^(†), —C(O)CH₂C(O)R^(†), —S(O)₂R^(†), —S(O)₂NR^(†) ₂,—C(S)NR^(†) ₂, —C(NH)NR^(†) ₂, or —N(R^(†))S(O)₂R^(†); wherein eachR^(†) is independently hydrogen, C₁₋₆ aliphatic which may be substitutedas defined below, unsubstituted —OPh, or an unsubstituted 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(†), taken together with their intervening atom(s) form anunsubstituted 3-12-membered saturated, partially unsaturated, or arylmono- or bicyclic ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R^(†) are independentlyhalogen, —R^(•), -(haloR^(•)), —OH, —OR^(•), —O(haloR^(•)), —CN,—C(O)OH, —C(O)OR^(•), —NH₂, —NHR^(•), —NR^(•) ₂, or —NO₂, wherein eachR^(•) is unsubstituted or where preceded by “halo” is substituted onlywith one or more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge etal., describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein byreference. Pharmaceutically acceptable salts of the compounds of thisinvention include those derived from suitable inorganic and organicacids and bases. Examples of pharmaceutically acceptable, nontoxic acidaddition salts are salts of an amino group formed with inorganic acidssuch as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuricacid and perchloric acid or with organic acids such as acetic acid,oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid ormalonic acid or by using other methods used in the art such as ionexchange. Other pharmaceutically acceptable salts include adipate,alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate,borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate,propionate, stearate, succinate, sulfate, tartrate, thiocyanate,p-toluenesulfonate, undecanoate, valerate salts, and the like.

Salts derived from appropriate bases include alkali metal, alkalineearth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. Representative alkali oralkaline earth metal salts include sodium, lithium, potassium, calcium,magnesium, and the like. Further pharmaceutically acceptable saltsinclude, when appropriate, nontoxic ammonium, quaternary ammonium, andamine cations formed using counterions such as halide, hydroxide,carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and arylsulfonate.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, Z and E double bond isomers,and Z and E conformational isomers. Therefore, single stereochemicalisomers as well as enantiomeric, diastereomeric, and geometric (orconformational) mixtures of the present compounds are within the scopeof the invention. Unless otherwise stated, all tautomeric forms of thecompounds of the invention are within the scope of the invention.Additionally, unless otherwise stated, structures depicted herein arealso meant to include compounds that differ only in the presence of oneor more isotopically enriched atoms. For example, compounds having thepresent structures including the replacement of hydrogen by deuterium ortritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enriched carbonare within the scope of this invention. Such compounds are useful, forexample, as analytical tools, as probes in biological assays, or astherapeutic agents in accordance with the present invention. In certainembodiments, a warhead moiety, R¹, of a provided compound comprises oneor more deuterium atoms.

3. DESCRIPTION OF EXEMPLARY EMBODIMENTS

In certain embodiments, the present invention provides inhibitors ofCaMKII. In some embodiments, such compounds include those of formula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   one of R^(2′) and R^(3′) is -L¹-R^(x), and the other is hydrogen;-   L¹ is a covalent bond or a straight or branched C₁₋₆ aliphatic    group, wherein one or more methylene groups are independently and    optionally replaced by —NR^(a)— or —O—;-   R^(x) is selected from the group consisting of NH₂, guanidino, 4-7    membered optionally substituted saturated heterocyclic ring having    1-2 heteroatoms independently selected from nitrogen, oxygen or    sulfur, and 5-6 membered heteroaromatic ring having 1-2 heteroatoms    independently selected from sulfur, nitrogen and oxygen;-   each of R³, R⁴, R⁵, R⁶, R⁷, R⁸, R^(4′), R^(5′), and R^(6′) is    independently selected from the group consisting of hydrogen,    halogen, —CN, —CF₃, —OR, —NR₂, —NO₂, —COOR, —CONR₂, and —R;-   each R^(a) is independently hydrogen or C₁₋₃ aliphatic; and-   each R is independently hydrogen or optionally substituted C₁₋₆    aliphatic.

As defined generally above, one of R^(2′) and R^(3′) is -L¹-R^(x), andthe other is hydrogen. In some embodiments R^(2′) is hydrogen and R^(3′)is -L¹-R^(x). In some embodiments one of R^(2′) is -L¹-R^(x) and R^(3′)is hydrogen.

As defined generally above, L¹ is a covalent bond or a straight orbranched C₁₋₆ aliphatic group, wherein one or more methylene groups areindependently and optionally replaced by —NR^(a)— or —O—. In someembodiments L¹ is a covalent bond. In some embodiments L¹ is a straightor branched C₁₋₆ aliphatic group wherein one or more methylene groupsare independently and optionally replaced by —NR^(a)— or —O—. In someembodiments L¹ is a C₁₋₃ aliphatic group. In some embodiments L¹ ismethylene. In some embodiments L¹ is a C₁₋₄ aliphatic group wherein onemethylene group is replaced by —NR^(a)— or —O—. In some embodiments, L¹is a C₁₋₄ alkylene group wherein the first methylene is replaced by—NR^(a)— (i.e. —NR^(a)—(CH₂)₀₋₃—). In some embodiments L¹ is a C₁₋₄alkylene group wherein the first methylene is replaced by —O— (i.e.—O—(CH₂)₀₋₃—).

As defined generally above, R^(x) is selected from the group consistingof NH₂, guanidino, 4-7 membered optionally substituted saturatedheterocyclic ring having 1-2 heteroatoms independently selected fromnitrogen or sulfur, and 5-6 membered heteroaromatic ring having 1-2heteroatoms independently selected from nitrogen and oxygen. In someembodiments R^(x) is NH₂. In some embodiments R^(x) is guanidine. Insome embodiments R^(x) is a 4-7 membered optionally substitutedsaturated heterocyclic group having 1-2 heteroatoms independentlyselected from nitrogen and oxygen. In some embodiments R^(x) ispiperazino. In some embodiments R^(x) is a 5-6 membered heteroaromaticring having 1-2 heteroatoms independently selected from nitrogen andoxygen. In some embodiments, R^(x) is imidazolo.

As defined generally above, each of R³, R⁴, R⁵, R⁶, R⁷, R⁸, R^(4′),R^(5′), and R^(6′) is independently selected from the group consistingof hydrogen, halogen, —CN, —CF₃, —OR, —NR₂, —NO₂, —COOR, —CONR₂, and —R.In some embodiments both of R⁶ and R⁷ are hydrogen. In some embodimentsR⁶ is hydrogen and R⁷ is selected from the group consisting of halogen,—CN, —CF₃, —OR, —NR₂, —NO₂, —COOR, —CONR₂, and —R. In some embodimentsR⁷ is hydrogen and R⁶ is selected from the group consisting of halogen,—CN, —CF₃, —OR, —NR₂, —NO₂, —COOR, —CONR₂, and —R. In some embodimentsboth R⁶ and R⁷ are independently selected from the group consisting ofhalogen, —CN, —CF₃, —OR, —NR₂, —NO₂, —COOR, —CONR₂, and —R. In someembodiments R⁶ is halogen. In some embodiments R⁶ is —CN. In someembodiments R⁶ is —OR. In some embodiments R⁶ is —OH. In someembodiments R⁶ is —NR₂. In some embodiments R⁶ is —NO₂. In someembodiments R⁶ is —COOR. In some embodiments R⁶ is —CONR₂. In someembodiments R⁶ is methoxy. In some embodiments R⁶ is —R, wherein R isC₁₋₃ aliphatic optionally substituted by one or more fluorines. In someembodiments R⁶ is methyl. In some embodiments R⁶ is trifluoromethyl. Insome embodiments R⁷ is halogen. In some embodiments R⁷ is —CN. In someembodiments R⁷ is —OR. In some embodiments R⁷ is —OH. In someembodiments R⁷ is —NR₂. In some embodiments R⁷ is —NO₂. In someembodiments R⁷ is —COOR. In some embodiments R⁷ is —CONR₂. In someembodiments R⁷ is methoxy. In some embodiments R⁷ is —R, wherein R isC₁₋₃ aliphatic optionally substituted by one or more fluorines. In someembodiments R⁷ is methyl. In some embodiments R⁷ is trifluoromethyl.

As defined generally above, each R^(a) is independently hydrogen or C₁₋₃aliphatic. In some embodiments each R^(a) is hydrogen. In someembodiments each R^(a) is C₁₋₃ aliphatic.

In certain embodiments, the present invention provides a compound offormula I wherein R³, R⁴, R⁵, R⁸, R^(4′), R^(5′), and R^(6′) are eachhydrogen, thereby forming a compound of formula I-a:

or a pharmaceutically acceptable salt thereof, wherein each of R^(2′),R^(3′), R⁶, and R⁷ is defined above and described in embodiments herein,both singly and in combination.

In certain embodiments, the present invention provides a compound offormula I wherein R^(2′) is hydrogen and R^(3′) is -L¹-R^(x), therebyforming a compound of formula II:

or a pharmaceutically acceptable salt thereof, wherein each of L¹,R^(x), R⁶, and R⁷ is defined above and described in embodiments herein,both singly and in combination.

In certain embodiments, the present invention provides a compound offormula I wherein R^(2′) is -L¹-R^(x) and R^(3′) is hydrogen, therebyforming a compound of formula III:

or a pharmaceutically acceptable salt thereof, wherein each of L¹,R^(x), R⁶, and R⁷ is defined above and described in embodiments herein,both singly and in combination.

In certain embodiments, the present invention provides a compound offormula II wherein L¹ is —NR^(a)—(CH₂)₀₋₃—, thereby forming a compoundof formula IV:

or a pharmaceutically acceptable salt thereof, wherein each of R^(x),R⁶, R⁷, and R^(a) is defined above and described in embodiments herein,both singly and in combination.

In certain embodiments, the present invention provides a compound offormula III wherein L¹ is —NR^(a)—(CH₂)₀₋₃—, thereby forming a compoundof formula V:

or a pharmaceutically acceptable salt thereof, wherein each of R^(x),R⁶, R⁷, and R^(a) is defined above and described in embodiments herein,both singly and in combination.

In certain embodiments, the present invention provides a compound offormula II wherein L¹ is —O—(CH₂)₀₋₃—, thereby forming a compound offormula VI:

or a pharmaceutically acceptable salt thereof, wherein each of R^(x),R⁶, and R⁷ is defined above and described in embodiments herein, bothsingly and in combination.

In certain embodiments, the present invention provides a compound offormula III wherein L¹ is —O—(CH₂)₀₋₃—, thereby forming a compound offormula VII:

or a pharmaceutically acceptable salt thereof, wherein each of R^(x),R⁶, and R⁷ is defined above and described in embodiments herein, bothsingly and in combination.

In certain embodiments, the present invention provides a compound offormula II wherein L¹ is a covalent bond, thereby forming a compound offormula VIII:

or a pharmaceutically acceptable salt thereof, wherein each of R^(x),R⁶, and R⁷ is defined above and described in embodiments herein, bothsingly and in combination.

In certain embodiments, the present invention provides a compound offormula III wherein L¹ is a covalent bond, thereby forming a compound offormula IX:

or a pharmaceutically acceptable salt thereof, wherein each of R^(x),R⁶, and R⁷ is defined above and described in embodiments herein, bothsingly and in combination.

Exemplary compounds of formula I are set forth in Table 1, below:

TABLE 1 Exemplary Compounds of Formula I Compound ID Compound StructureI-1 

I-2 

I-3 

I-4 

I-5 

I-6 

I-7 

I-8 

I-9 

I-10

I-11

I-12

I-13

I-14

I-15

I-16

In certain embodiments, the present invention provides any compoundselected from those depicted in Table 1, above, or a pharmaceuticallyacceptable salt thereof.

Compounds or salts thereof provided by the present invention may beutilized in any of a variety of forms. For example, in some embodiments,provided compounds (or salts thereof) are utilized in a solid form; insome such embodiments, provided compounds (or salts thereof) areutilized in an amorphous solid form. In some embodiments, providedcompounds are utilized in a crystalline solid form. In some embodiments,provided compounds (or salts thereof) are utilized in a solid form(e.g., a crystalline solid form) that is a solvate or hydrate.

4. USES, FORMULATION AND ADMINISTRATION AND PHARMACEUTICALLY ACCEPTABLECOMPOSITIONS

According to some embodiments, the invention provides a compositioncomprising a compound of this invention or a pharmaceutically acceptablederivative thereof and a pharmaceutically acceptable carrier, adjuvant,or vehicle.

In certain embodiments, the invention provides compositions containingan amount of compound effective to measurably inhibit CaMKII, in abiological sample or in a patient. In certain embodiments, the amount ofcompound in compositions of this invention is such that is effective tomeasurably inhibit a CaMKII mediated biological process in a biologicalsample or in a patient. In certain embodiments, provided compositionscontain a unit dose amount of a compound described herein, whereinadministration of such unit dose amount as part of a therapeutic regimencorrelates with a desired pharmacologic and/or therapeutic outcome.

In certain embodiments, a composition of this invention is formulatedfor administration to a patient in need of such composition. In someembodiments, a composition of this invention is formulated for oraladministration to a patient.

As used herein, a “dosing regimen” or “therapeutic regimen” refers to aset of unit doses (typically more than one) that are administeredindividually to a subject, typically separated by periods of time. Insome embodiments, a given therapeutic agent has a recommended dosingregimen, which may involve one or more doses. In some embodiments, adosing regimen comprises a plurality of doses each of which areseparated from one another by a time period of the same length; in someembodiments, a dosing regime comprises a plurality of doses and at leasttwo different time periods separating individual doses. In someembodiments, all doses within a dosing regimen are of the same unit doseamount. In some embodiments, different doses within a dosing regimen areof different amounts. In some embodiments, a dosing regimen comprises afirst dose in a first dose amount, followed by one or more additionaldoses in a second dose amount different from the first dose amount. Insome embodiments, a dosing regimen comprises a first dose in a firstdose amount, followed by one or more additional doses in a second doseamount same as the first dose amount.

The term “patient,” as used herein, means an animal, often a mammal, andin many embodiments a human.

The term “pharmaceutically acceptable carrier, adjuvant, or vehicle”, asused herein, refers to a non-toxic carrier, adjuvant, or vehicle thatdoes not destroy the pharmacological activity of the compound with whichit is formulated. Pharmaceutically acceptable carriers, adjuvants orvehicles that may be used in the compositions of this invention include,but are not limited to, ion exchangers, alumina, aluminum stearate,lecithin, serum proteins, such as human serum albumin, buffer substancessuch as phosphates, glycine, sorbic acid, potassium sorbate, partialglyceride mixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, sodium carboxymethylcellulose,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,polyethylene glycol and wool fat.

A “pharmaceutically acceptable derivative” means any non-toxic salt,ester, or salt of an ester of a compound of this invention that, uponadministration to a recipient, is capable of providing, either directlyor indirectly, a compound of this invention or an inhibitorily activemetabolite or residue thereof.

As used herein, the term “inhibitorily active metabolite or residuethereof” means that a metabolite or residue thereof is also an inhibitorof CaMKII or is retains therapeutic activity in treating the samedisease, disorder or condition.

Compositions of the present invention may be formulated for anyappropriate route of administration. For example, in some embodiments,provided compositions may be administered orally, parenterally, byinhalation spray, topically, rectally, nasally, buccally, vaginally orvia an implanted reservoir. The term “parenteral” as used hereinincludes subcutaneous, intravenous, intramuscular, intra-articular,intra-synovial, intrasternal, intrathecal, intrahepatic, intralesionaland intracranial injection or infusion techniques. In some embodiments,provided compositions are administered orally, intraperitoneally orintravenously. Sterile injectable forms of the compositions of thisinvention may be aqueous or oleaginous suspension. Such suspensions maybe formulated according to techniques known in the art using suitabledispersing or wetting agents and suspending agents.

In some embodiments, pharmaceutically acceptable compositions of theinvention may be formulated as injectable preparations. Injectablepreparations, for example, sterile injectable aqueous or oleaginoussuspensions may be formulated according to the known art using suitabledispersing or wetting agents and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution,suspension or emulsion in a nontoxic parenterally acceptable diluent orsolvent, for example, as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

In some embodiments, injectable formulations can be sterilized, forexample, by filtration through a bacterial-retaining filter, or byincorporating sterilizing agents in the form of sterile solidcompositions which can be dissolved or dispersed in sterile water orother sterile injectable medium prior to use.

In some embodiments, for example in order to prolong effects of acompound or composition, it may be desirable to slow the absorption ofthe compound from subcutaneous or intramuscular injection. This may beaccomplished by the use of a liquid suspension of crystalline oramorphous material with poor water solubility. The rate of absorption ofthe compound then depends upon its rate of dissolution that, in turn,may depend upon crystal size and crystalline form. Alternatively oradditionally, delayed absorption of a parenterally administered compoundform is accomplished by dissolving or suspending the compound in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the compound in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of compound topolymer and the nature of the particular polymer employed, the rate ofcompound release can be controlled. Examples of other biodegradablepolymers include poly(orthoesters) and poly(anhydrides). Depotinjectable formulations are also prepared by entrapping the compound inliposomes or microemulsions that are compatible with body tissues.

In some embodiments, sterile injectable preparations may be or include asterile injectable solution or suspension in a non-toxic parenterallyacceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or di-glycerides. Fatty acids, such as oleic acid andits glyceride derivatives are useful in the preparation of injectables,as are natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. Such oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, such as carboxymethyl cellulose or similar dispersingagents that are commonly used in the formulation of pharmaceuticallyacceptable dosage forms including emulsions and suspensions. Othercommonly used surfactants, such as Tweens, Spans and other emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms may also be used for the purposes of formulation.

Pharmaceutically acceptable compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include lactose andcorn starch. Lubricating agents, such as magnesium stearate, are alsotypically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like.

In some embodiments, provided compounds can be in micro-encapsulatedform with one or more excipients as noted above. Solid dosage forms suchas tablets, dragees, capsules, pills, and granules can be prepared withcoatings and shells such as enteric coatings, release controllingcoatings and other coatings well known in the pharmaceutical formulatingart. In such solid dosage forms, the active compound may be admixed withat least one inert diluent such as sucrose, lactose or starch. Suchdosage forms may also comprise, as is normal practice, additionalsubstances other than inert diluents, e.g., tableting lubricants andother tableting aids such a magnesium stearate and microcrystallinecellulose. In the case of capsules, tablets and pills, the dosage formsmay also comprise buffering agents. They may optionally containopacifying agents and can also be of a composition that they release theactive ingredient(s) only, or preferentially, in a certain part of theintestinal tract, optionally, in a delayed manner. Examples of embeddingcompositions that can be used include polymeric substances and waxes.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Alternatively or additionally, pharmaceutically acceptable compositionsof this invention may be administered in the form of suppositories forrectal administration. Such compositions can be prepared by combining aprovided compound with a suitable non-irritating excipient that is solidat room temperature but liquid at rectal temperature and therefore willmelt in the rectum to release the drug. Such materials include cocoabutter, beeswax and polyethylene glycols.

In some embodiments, pharmaceutically acceptable compositions of thisinvention may be administered topically, especially when the target oftreatment includes areas or organs readily accessible by topicalapplication, including diseases of the eye, the skin, or the lowerintestinal tract. Suitable topical formulations are readily prepared foreach of these areas or organs.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, provided pharmaceutically acceptablecompositions may be formulated in a suitable ointment containing theactive component suspended or dissolved in one or more carriers.Carriers for topical administration of compounds of this inventioninclude, but are not limited to, mineral oil, liquid petrolatum, whitepetrolatum, propylene glycol, polyoxyethylene, polyoxypropylenecompound, emulsifying wax and water. Alternatively or additionally,provided pharmaceutically acceptable compositions can be formulated in asuitable lotion or cream containing the active components suspended ordissolved in one or more pharmaceutically acceptable carriers. Suitablecarriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, and eye drops are also contemplatedas being within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

For ophthalmic use, provided pharmaceutically acceptable compositionsmay be formulated as micronized suspensions in isotonic, pH adjustedsterile saline, or, preferably, as solutions in isotonic, pH adjustedsterile saline, either with or without a preservative such asbenzylalkonium chloride. Alternatively or additionally, for ophthalmicuses, the pharmaceutically acceptable compositions may be formulated inan ointment such as petrolatum.

In some embodiments, pharmaceutically acceptable compositions of thisinvention may be administered by nasal aerosol or inhalation. Suchcompositions may be prepared according to techniques well-known in theart of pharmaceutical formulation, for example as solutions in saline,employing benzyl alcohol or other suitable preservatives, absorptionpromoters to enhance bioavailability, fluorocarbons, and/or otherconventional solubilizing or dispersing agents.

In some embodiments, pharmaceutically acceptable compositions of thisinvention are formulated for oral administration. Such formulations maybe administered with or without food. In some embodiments,pharmaceutically acceptable compositions of this invention areadministered without food. In some embodiments, pharmaceuticallyacceptable compositions of this invention are administered with food.

The amount of compounds of the present invention that may be combinedwith the carrier materials to produce a composition in a single dosageform will vary depending upon the host treated, the particular mode ofadministration. In some embodiments provided compositions are formulatedso that a dosage of between 0.01-100 mg/kg body weight/day of theinhibitor can be administered to a patient receiving these compositions.

It should also be understood that a specific dosage and treatmentregimen for any particular patient may depend upon a variety of factors,including the activity of the specific compound employed, the age, bodyweight, general health, sex, diet, time of administration, rate ofexcretion, drug combination, and the judgment of the treating physicianand the severity of the particular disease being treated. In someembodiments, amount of a compound of the present invention included in acomposition described herein is determined by activity and/orbioavailability of the particular compound, so that compositions ofdifferent compounds may include different absolute amounts of compound.

Uses of Compounds and Pharmaceutically Acceptable Compositions

Compounds and compositions described herein are useful in the treatmentof any of a variety of diseases, disorders, and conditions. In someembodiments, provided compounds and compositions are useful in thetreatment of diseases, disorders, or conditions associated with activityof CaMKII.

Ca²⁺/calmodulin-dependent protein kinase (CaMKII) is a serine/threoninekinase. Several lines of evidence strongly support the concept of directinhibition of CaMKII activity as an important therapeutic target fortreating a range of diseases including cardiovascular diseases such asatrial fibrillation, ventricular arrhythmia, heart failure, cardiachypertrophy, atherosclerosis, and in-stent restenosis in coronary arterydisease; use in cardioprotection; inflammatory lung diseases such asasthma; neurological diseases and conditions such as pain, stroke,ischemia, hypoxia, opioid tolerance and dependence, and maculardegeneration; metabolic disorders such as type II diabetes, insulinresistance, and obesity; cancer and other proliferative disorders, suchas osteosarcoma, melanoma, and prostate cancer; bone diseases such asosteoporosis; and inflammatory diseases such as rheumatoid arthritis.

Ca²⁺ acts as an informational or second messenger to modulate theeffects of many hormones and neurotransmitters as well as duringdepolarization of excitable membranes in heart, skeletal muscle, andbrain. In many instances Ca²⁺ mediates its effects via a Ca²⁺ bindingprotein termed calmodulin. The Ca²⁺/calmodulin complex can coordinatemultiple cellular functions by activation CaMKII, a majormultifunctional protein kinase in diverse tissues, including heart,brain, skeletal muscle, and liver. Strong activation of the kinase byCa²⁺ can result in a self-phosphorylation or autophosphorylation thatmakes the kinase persistently active, a state that can also be achievedby oxidation.

CaMKII refers to four highly conserved isoforms of CaMKII, α, β, γ, andδ. CaMKII α, β, and γ are widely expressed, with the δ isoformspredominating in heart. Upon activation, CaMKII catalyzes a transfer ofphosphate from ATP onto specific sites on select substrate proteins,such as the Ryanodine receptor RyR2 and the L-type Ca²⁺ channel inheart. These isoforms have been crystallized as truncated monomericcatalytic domains (Rellos P, Pike A C W, Niesen F H, Salah E, Lee W H,von Delft F, Knapp S. (2010) “Structure of the CaMKIIdelta/CalmodulinComplex Reveals the Molecular Mechanism of CaMKII Kinase Activation”PLoS Biol 8:e1000426) as well as in the native multimeric holoenzyme(Chao L H, Stratton M M, Lee I-H, Rosenberg O S, Levitz J, Mandell D J,Kortemme T, Groves J T, Schulman H, Kuriyan J. (2011) “A Mechanism forTunable Autoinhibition in the Structure of a HumanCa²⁺/Calmodulin-Dependent Kinase II Holoenzyme” Cell 146:732-745).Comparison of the sequences, three-dimensional structure of thecatalytic sites, and limited pharmacology suggest a great deal ofhomology between the isoforms, and ATP competitive inhibitors of any ofthe isoforms will likely inhibit all of the others. ATP competitiveinhibitors are also known to block both the Ca²⁺/calmodulin-activatedenzyme as well the persistently activated forms generated byautophosphorylation or oxidation by reactive oxygen.

Pharmacological and genetic inhibition of CaMKII reduces ryanodinereceptor-mediated calcium leaks and blocked induction of atrialfibrillation in a mouse model of atrial fibrillation. In atrial cellsfrom atrial fibrillation patients CaMKII activity is increased leadingto calcium leaks that promote atrial fibrillation, while inhibition ofthe kinase reduces the calcium leak. (Dobrev D, et al., “Novel moleculartargets for atrial fibrillation therapy” (2012) Nature Reviews DrugDiscovery 11:275-291; Chelu M G, et al., “Calmodulin kinase II-mediatedsarcoplasmic reticulum Ca²⁺ leak promotes atrial fibrillation in mice”(2009) J Clin Invest 119:1940-1951; Neef S, et al., “CaMKII-dependentdiastolic SR Ca²⁺ leak and elevated diastolic Ca²⁺ levels in rightatrial myocardium of patients with atrial fibrillation” (2010) Circ Res106:1134-1144).

Pharmacological inhibition of CaMKII is shown to reduce cardiacarrhythmogenesis in vitro and in vivo, including inhibition of torsadesthat lead to sudden death. (Anderson M E, et al., “KN-93, an inhibitorof multifunctional Ca⁺⁺/Calmodulin-dependent protein kinase, decreasesearly afterdepolarizations in rabbit heart” (1998) J Pharmacol Exp Ther287:996-1006; Sag C M, et al., “Calcium/calmodulin-dependent proteinkinase II contributes to cardiac arrhythmogenesis in heart failure”(2009) Circ Heart Fail 2:664-675; Erickson J R, Anderson M E, “CaMKIIand Its role in cardiac arrhythmia” (2008) Journal of CardiovascularElectrophysiology 19:1332-1336). CaMKII integrates several proarrhythmicpathways that augment CaMKII activity via Ca²⁺ and reactive oxygenspecies and which, in turn, acts on the ryanodine receptor, thevoltage-dependent calcium channel (Cav1.2), and the Na channel (Nav1.5)to promote arrhythmia (Rokita A G and Anderson M E “New TherapeuticTargets in Cardiology Arrhythmias and Ca2+/Calmodulin-Dependent KinaseII (CaMKII)” (2012) Circulation 126:2125-2139).

Studies implicate CaMKII in heart failure and structural heart diseasein both mouse models and human heart tissue. Pharmacological andgenetic-based inhibition of CaMKII was shown to protect cellularmechanical function and preserves calcium homeostasis after myocardialinfarction. CaMKII is increased in cells from failing human heart cellsand its pharmacological inhibition improves contractility by anestablished CaMKII pathway. (Schulman H, Anderson M E,“Ca/Calmodulin-dependent Protein Kinase II in Heart Failure” (2010) DrugDiscovery Today: Disease Mechanisms 7:e117-e122; Zhang R, et al.,“Calmodulin kinase II inhibition protects against structural heartdisease” (2005) Nat Med 11:409-417; Sossalla S, et al., “Inhibition ofElevated Ca²⁺/Calmodulin-Dependent Protein Kinase II ImprovesContractility in Human Failing Myocardium” (2010) Circulation Research107:1150-1161).

Genetic activation and pharmacological inhibition of CaMKII were used toshow that CaMKII mediates cardiac hypertrophy while genetic deletion ofδ-CaMKII protected the heart from pathological cardiac hypertrophy andremodeling after pressure overload. (Backs J, et al., “The delta isoformof CaM kinase II is required for pathological cardiac hypertrophy andremodeling after pressure overload” (2009) Proc Natl Acad Sci USA106:2342-2347; Zhang T, et al., “The cardiac-specific nuclear delta(B)isoform of Ca²⁺/calmodulin-dependent protein kinase II induceshypertrophy and dilated cardiomyopathy associated with increased proteinphosphatase 2A activity” (2002) J Biol Chem 277:1261-1267; Anderson M E,et al., “CaMKII in myocardial hypertrophy and heart failure” (2011)Journal of Molecular and Cellular Cardiology 51:468-473).

CaMKII inhibition is found to be effective in several forms ofcardioprotection, including from cardiotoxicity caused by cancer therapy(doxorubicin), following heart attack or ischemia-reperfusion, e.g. forthe case of acute intervention for heart attack (primary angioplasty),and for patients with mutations leading to sudden death, such ascatecholaminergic polymorphic ventricular tachycardia. (Sag C M, et al.,“CaMKII-dependent SR Ca leak contributes to doxorubicin-induced impairedCa handling in isolated cardiac myocytes” (2011) Journal of Molecularand Cellular Cardiology 51:749-759; Zhang R, et al., “Calmodulin kinaseII inhibition protects against structural heart disease” (2005) Nat Med11:409-417; Liu N, et al., “Calmodulin kinase II inhibition preventsarrhythmias in RyR2(R4496C+/−) mice with catecholaminergic polymorphicventricular tachycardia” (2011) Journal of Molecular and CellularCardiology 50:214-222; Joiner, M-L A, et al., “CaMKII determinesmitochondrial stress responses in heart” (2012). Nat Med, DOI:10.1038/nature11444, published online Oct. 10, 2012).

Atherosclerosis pathology includes both the constriction of thevasculature as well as plaque disruption. CaMKII inhibition blocksproliferation of vascular cells as well as mediating ER stressors thatlead to apoptosis that underlie plaque disruption. (Timmins J M, et al.,“Calcium/calmodulin-dependent protein kinase II links ER stress with Fasand mitochondrial apoptosis pathways” (2009) The Journal of ClinicalInvestigation 119:2925-2941; Li W, et al., “The multifunctionalCa²⁺/calmodulin-dependent kinase IIδ (CaMKIIδ) controls neointimaformation after carotid ligation and vascular smooth muscle cellproliferation through cell cycle regulation by p21” (2011) J Biol Chem286:7990-7999).

Studies suggest that CaMKII is an important, but previously unrecognizedpro-asthmatic signal, linking the pro-oxidant environment of theasthmatic airways with downstream inflammatory and remodeling events.CaMKII activity in the epithelium may be required for enhancingeosinophilic recruitment to the lung, through a ROS-CaMKII-eotaxin-1dependent pathway. Inhibition of CaMKII activity may be a novel targetin future asthma therapies. (Sanders P N, et al., “Camkii As APro-Asthmatic Signal” (2011) Am J Respir Care Med 183:A2795, May 6, 2011poster presentation).

Smooth muscle proliferation contributes to vascular remodeling andobstructive vasculopathies such as atherosclerosis and restenosisfollowing percutaneous coronary interventions and inhibition of thekinase blocks vascular smooth muscle proliferation and neointimalformation that lead to restenosis. (Li W, et al., “The multifunctionalCa²⁺/calmodulin-dependent kinase IIδ (CaMKIIδ) controls neointimaformation after carotid ligation and vascular smooth muscle cellproliferation through cell cycle regulation by p21” (2011) J Biol Chem286:7990-7999; House S J, Singer H A, “CaMKII-delta isoform regulationof neointima formation after vascular injury” (2008) Arterioscler ThrombVasc Biol 28:441-447).

Pharmacological and genetic suppression of CaMKII has been used todemonstrate a reduction in central and peripheral pain due to injury orinflammation as well as in sensitization to pain. (Zeitz K P, et al.,“The contribution of autophosphorylated alpha-calcium-calmodulin kinaseII to injury-induced persistent pain” (2004) Neuroscience 128:889-898;Luo F, et al., “Reversal of chronic inflammatory pain by acuteinhibition of Ca²⁺/calmodulin-dependent protein kinase II” (2008) JPharmacol Exp Ther 325:267-275; Chen Y, et al.,“Ca²⁺/Calmodulin-dependent protein kinase IIα is required for theinitiation and maintenance of opioid-induced hyperalgesia” (2010) JNeurosci 30:38-46; Crown E D, et al., “Calcium/calmodulin dependentkinase II contributes to persistent central neuropathic pain followingspinal cord injury” (2012) Pain 153:710-721).

Inhibition of CaMKII is neuroprotective, reducing damage due to hypoxiain stroke models. Reduction of atrial fibrillation by Inhibition ofCaMKII would also reduce stroke incidence. (Vest R S, et al., “Effectivepost-insult neuroprotection by a novel CaMKII inhibitor” (2010) J BiolChem 285:20675-20682; Ashpole N M, et al., “Calcium/Calmodulin-dependentProtein Kinase II (CaMKII) Inhibition Induces Neurotoxicity viaDysregulation of Glutamate/Calcium Signaling and Hyperexcitability”(2012) Journal of Biological Chemistry 287:8495-8506; Dobrev D, et al.,“Novel molecular targets for atrial fibrillation therapy” (2012) NatureReviews Drug Discovery 11:275-291).

Stimulation of opiate receptors increases CaMKII and leads to toleranceand dependence that are reduced by inhibition of CaMKII. (Liang D, etal., “Increased expression of Ca2+/calmodulin-dependent protein kinaseII alpha during chronic morphine exposure” (2004) Neuroscience123:769-775; Fan G H, et al., “Inhibition ofcalcium/calmodulin-dependent protein kinase II in rat hippocampusattenuates morphine tolerance and dependence” (1999) Mol Pharmacol56:39-45).

Inhibition of CaMKII reduces the VEGF pathway that mediates increasedvascularization or angiogenesis of retinal endothelial cells. (BanumathiE, et al., “VEGF-induced retinal angiogenic signaling is criticallydependent on Ca²⁺ signaling via Ca²⁺/calmodulin-dependent protein kinaseII” (2011) Investigative Ophthalmology & Visual Science 52:3103-3111).

CaMKII may have several sites of action that support the notion ofCaMKII inhibition in type II diabetes. CaMKII modulates insulinsignaling that suggest a role in the pathogenesis of insulin resistance.In liver, CaMKII regulates glucose production and its inhibition wouldbe beneficial in diabetes. (Illario M, et al.,“Calcium-calmodulin-dependent kinase II (CaMKII) mediatesinsulin-stimulated proliferation and glucose uptake” (2009) CellularSignaling 21:786-792; Ozcan L, et al., “Calcium Signaling through CaMKIIRegulates Hepatic Glucose Production in Fasting and Obesity” (2012) CellMetabolism 15:739-751).

Studies show that pharmacological inhibition of CaMKII reducesproliferation of osteosarcoma cell lines and indicates changes in signaltransduction related to growth. The inhibitor administered to mice witha human osteosarcoma xenograft markedly decreases tumor size. (Yuan K,et al., “α-CaMKII controls the growth of human osteosarcoma byregulating cell cycle progression” (2007) Lab Invest 87:938-950).

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) providesa pathway in melanoma therapy but melanoma is often resistant to TRAILafter metastasis. Inhibition of CaMKII signaling by use of a dominantnegative form of the kinase was shown to restore the sensitivity ofmelanoma to cell death via TRAIL. (Xiao C, et al., “Inhibition ofCaMKII-mediated c-FLIP expression sensitizes malignant melanoma cells toTRAIL-induced apoptosis” (2005) Exp Cell Res 304:244-255).

Studies have shown that proliferation and invasion of prostate cancercell lines is reduced by pharmacological inhibition of CaMKII. Kinaseinhibition was used to show that it is important for prostate cancercell survival and promotes their progression to an androgen-independentstate. (Mamaeva O A, et al., “Calcium/calmodulin-dependent kinase IIregulates notch-1 signaling in prostate cancer cells” (2009) J CellBiochem 106:25-32; Rokhlin O W, et al., “Calcium/calmodulin-dependentkinase II plays an important role in prostate cancer cell survival”(2007) Cancer Biol Ther 6:732-742).

Pharmacological inhibition of CaMKII reduces differentiation ofosteoclasts and suppresses bone resorption characteristic ofosteoporosis. (Ang E S M, et al., “Calcium/calmodulin-dependent kinaseactivity is required for efficient induction of osteoclastdifferentiation and bone resorption by receptor activator of nuclearfactor kappa B ligand (RANKL)” (2007) Journal of cellular physiology212:787-795).

Pharmacological and genetic suppression of CaMKII demonstrated its rolein the production of proinflammatory cytokines and interferon inmacrophages. A small molecule inhibitor of CaMKII was used to show thatit required for tumour necrosis factor-related apoptosis inducing ligand(TRAIL)-mediated apoptosis of fibroblast-like synovial cells, suggestingthat it is a target for rheumatoid arthritis therapy. (Liu X, et al.,“CaMKII promotes TLR-triggered proinflammatory cytokine and type Iinterferon production by directly binding and activating TAK1 and IRF3in macrophages” (2008) Blood 112:4961-4970; Fujikawa K, et al.,“Calcium/calmodulin-dependent protein kinase II (CaMKII) regulatestumour necrosis factor-related apoptosis inducing ligand(TRAIL)-mediated apoptosis of fibroblast-like synovial cells (FLS) byphosphorylation of Akt” (2009) Clinical and experimental rheumatology27:952-957).

A genetic model of CaMKII inhibition as well as a genetic knockout ofthe CaMKIIdelta show that the heart is protected from fibrosis andstructural remodeling normally induced by myocardial infarction and bypressure overload, respectively (Zhang R, et al. “Calmodulin kinase IIinhibition protects against structural heart disease” (2005) Nat Med11:409-417; Backs J, et al. “The delta isoform of CaM kinase II isrequired for pathological cardiac hypertrophy and remodeling afterpressure overload” (2009) Proc Natl Acad Sci USA 106:2342-2347). Theseconclusions are consistent with the observation that pharmacologicalinhibition of CaMKII reduces fibroblast proliferation, secretion offibroblast growth factor β1, and decreased expression of severalmetalloproteinases that underlie cardiac hypertrophy and fibrosis (ZhangW, et al. “Inhibition of calcium-calmodulin-dependent kinase IIsuppresses cardiac fibroblast proliferation and extracellular matrixsecretion” (2009) J Cardiovascular Pharmacology 55:96-105). Fibrosis isalso promoted by activation of CaMKII via reactive oxygen, such asduring Angiotensin II stimulation. These data support an indication forCaMKII inhibitors in reducing fibrosis in a number of possibleindications in which fibrosis is prominent, including myocardialfibrosis, cardiac hypertrophy, and myocardial infarction.

The activity of a compound utilized in this invention as an inhibitor ofCaMKII or treatment for a CaMKII-mediated disease, disorder orcondition, may be assayed in vitro or in vivo. An in vivo assessment ofthe efficacy of the compounds of the invention may be made using ananimal model of a CaMKII-mediated disease, disorder or condition, e.g.,a rodent or primate model. Cell-based assays may be performed using,e.g., a cell line isolated from a tissue that expresses CaMKII.Additionally, biochemical or mechanism-based assays, e.g., transcriptionassays using a purified protein, Northern blot, RT-PCR, etc., may beperformed. In vitro assays include assays that determine cellmorphology, protein expression, and/or the cytotoxicity, enzymeinhibitory activity, and/or the subsequent functional consequences oftreatment of cells with compounds of the invention. Alternate oradditional in vitro assays may be used to quantitate the ability of theinhibitor to bind to protein or nucleic acid molecules within the cell.Inhibitor binding may be measured by radiolabelling the inhibitor priorto binding, isolating the inhibitor/target molecule complex anddetermining the amount of radiolabel bound. Alternatively oradditionally, inhibitor binding may be determined by running acompetition experiment where new inhibitors are incubated with purifiedproteins or nucleic acids bound to known radioligands. Detailedconditions of exemplary systems for assaying a compound utilized in thisinvention as an inhibitor of CaMKII are set forth in the Examples below.Such assays are exemplary and not intended to limit the scope of theinvention. The skilled practitioner can appreciate that modificationscan be made to conventional assays to develop equivalent or other assaysthat can be employed to comparably assess activity or otherwisecharacterize compounds and/or compositions as described herein.

As used herein, the terms “treatment,” “treat,” and “treating” refer toreversing, alleviating, delaying the onset of, reducing incidence orseverity, or inhibiting the progress of a disease, disorder orcondition, or one or more symptoms thereof, as described herein. In someembodiments, treatment may be administered after one or more symptomshave developed. In other embodiments, treatment may be administered inthe absence of symptoms. For example, treatment may be administered to asusceptible individual prior to the onset of symptoms (e.g., in light ofa history of symptoms and/or in light of genetic or other susceptibilityfactors). Treatment may also be continued after symptoms have resolved,for example to prevent or delay their recurrence.

Compounds and/or compositions described herein may be administered usingany amount and any route of administration effective for treating adisease, disorder, or condition. In some embodiments, compounds and/orcompositions are administered in an amount and/or by a route effectivefor treating a cardiovascular disease, disorder or condition, aninflammatory disease, disorder or condition, a neurological disease,disorder or condition, an ocular disease, disorder or condition, ametabolic disease, disorder or condition, a cancer or otherproliferative disease, disorder or condition, a bone disease, disorderor condition, or an addictive disease, disorder, or condition.

In some embodiments, compounds and/or compositions described herein maybe administered using any amount and any route of administrationeffective for treating or lessening the severity of a disease, disorderor condition associated with CaMKII.

In some embodiments, compounds and/or compositions may be administeredusing any amount and any route of administration effective for treatinga cardiovascular disease, disorder, or condition. In some embodiments,the cardiovascular disease, disorder or condition is a disease of theheart. In some embodiments, the cardiovascular disease, disorder orcondition is a disease of the vasculature. In some embodiments, thecardiovascular disease, disorder or condition is selected from atrialfibrillation, ventricular arrhythmia, heart failure, cardiachypertrophy, atherosclerosis, fibrosis, or restenosis. In someembodiments the restenosis is in-stent restenosis in coronary arterydisease.

In some embodiments, provided compounds and/or compositions may beadministered using any amount and any route of administration effectivefor achieving cardioprotection from cardiotoxicity. In some embodiments,the cardiotoxicity avoided by administration of the compounds andcompositions of the invention is due to drug therapy, heart attack,ischemia-reperfusion injury, or mutations leading to sudden death suchas catecholaminergic polymorphic ventricular tachycardia.

In some embodiments, the compounds and compositions, according to themethod of the present invention, may be administered using any amountand any route of administration effective for treating an inflammatorydisease, disorder or condition. In some embodiments, the inflammatorydisease, disorder or condition is asthma or rheumatoid arthritis.

In some embodiments, the compounds and compositions, according to themethod of the present invention, may be administered using any amountand any route of administration effective for treating a neurologicaldisease, disorder or condition. In some embodiments, the neurologicaldisease, disorder or condition is pain or stroke.

In some embodiments, the compounds and compositions, according to themethod of the present invention, may be administered using any amountand any route of administration effective for treating an addictivedisease, disorder or condition. In some embodiments the addictivedisease, disorder, or condition is opioid tolerance or dependence.

In some embodiments, the compounds and compositions, according to themethod of the present invention, may be administered using any amountand any route of administration effective for treating an oculardisease, disorder or condition. In some embodiments, the ocular disease,disorder or condition is macular degeneration.

In some embodiments, the compounds and compositions, according to themethod of the present invention, may be administered using any amountand any route of administration effective for treating a metabolicdisease, disorder or condition. In some embodiments, the metabolicdisease, disorder or condition is diabetes. In some embodiments, thediabetes is type II diabetes.

In some embodiments, the compounds and compositions, according to themethod of the present invention, may be administered using any amountand any route of administration effective for treating a cancer oranother proliferative disease, disorder or condition. In someembodiments, the cancer or other proliferative disease, disorder orcondition is an osteosarcoma, a melanoma, or a prostate cancer.

In some embodiments, the compounds and compositions, according to themethod of the present invention, may be administered using any amountand any route of administration effective for treating a bone disease,disorder or condition. In some embodiments, the bone disease, disorderor condition is osteoporosis.

In will be appreciated by those skilled in the art that the exact amountof a provided compound or composition may vary from subject to subject,depending on the species, age, and general condition of the subject, theseverity of the infection, the particular agent, its mode ofadministration, and the like.

In some embodiments, compounds of the invention are formulated in dosageunit form, for example for ease of administration and uniformity ofdosage. The expression “dosage unit form” or “unit dosage” as usedherein refers to a physically discrete unit of agent appropriate for thepatient to be treated. It will be understood, however, that total dailyusage of the compounds and compositions of the present invention may bedecided by the attending physician within the scope of sound medicaljudgment. The specific effective dose level for any particular patientor organism may depend upon a variety of factors including the disorderbeing treated and the severity of the disorder; the activity of thespecific compound employed; the specific composition employed; the age,body weight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts.

According to some embodiments, the invention relates to a method ofinhibiting CaMKII in a biological sample comprising the step ofcontacting said biological sample with a compound of this invention, ora composition comprising said compound.

The term “biological sample”, as used herein, includes, withoutlimitation, cell cultures or extracts thereof; biopsied materialobtained from a mammal or extracts thereof; and blood, saliva, urine,feces, semen, tears, or other body fluids or extracts thereof.

Inhibition of enzymes in a biological sample is useful for a variety ofpurposes that are known to one of skill in the art. Examples of suchpurposes include, but are not limited to biological assays, geneexpression studies, and biological target identification.

Some embodiments of the present invention relate to a method ofinhibiting CaMKII in a patient comprising the step of administering tosaid patient a compound of the present invention, or a compositioncomprising said compound.

In some embodiments, the invention relates to a method of inhibitingCaMKII activity in a patient comprising the step of administering tosaid patient a compound of the present invention, or a compositioncomprising said compound. In certain embodiments, the present inventionprovides a method for treating a disease, disorder or condition mediatedby CaMKII, in a patient in need thereof, comprising the step ofadministering to said patient a compound according to the presentinvention or pharmaceutically acceptable composition thereof. Suchdiseases, disorders and conditions are described in detail herein.

In some embodiments compounds and/or compositions of the presentinvention may be used in a method of treating a cardiovascular disease,disorder, or condition, an inflammatory disease, disorder or condition,a neurological disease, disorder or condition, an ocular disease,disorder or condition, a metabolic disease, disorder or condition, acancer or other proliferative disease, disorder or condition, or a bonedisease, disorder or condition. In certain embodiments the compounds andcompositions of the present invention may be used to treat acardiovascular disease, disorder or condition, an inflammatory disease,disorder or condition, a neurological disease, disorder or condition, anocular disease, disorder or condition, a metabolic disease, disorder orcondition, a cancer or other proliferative disease, disorder orcondition, or a bone disease, disorder or condition in a mammal. Incertain embodiments the mammal is a human patient.

In some embodiments the present invention provides a method of treatinga cardiovascular disease, disorder or condition, an inflammatorydisease, disorder or condition, a neurological disease, disorder orcondition, an ocular disease, disorder or condition, a metabolicdisease, disorder or condition, a cancer or other proliferative disease,disorder or condition, or a bone disease, disorder or condition,comprising administering a compound or composition of the presentinvention to a patient in need thereof. In certain embodiments themethod of treating a cardiovascular disease, disorder or condition, aninflammatory disease, disorder or condition, a neurological disease,disorder or condition, an ocular disease, disorder or condition, ametabolic disease, disorder or condition, a cancer or otherproliferative disease, disorder or condition, or a bone disease,disorder or condition comprises administering compounds and compositionsof the present invention to a mammal. In certain embodiments the mammalis a human.

In certain embodiments, the present invention provides a method oftreating a cardiovascular disease, disorder or condition, comprisingadministering a compound or composition of the present invention to apatient with a cardiovascular disease, disorder or condition. In certainembodiments, the method of treating a cardiovascular disease, disorderor condition comprises administering compounds and compositions of thepresent invention to a mammal. In certain embodiments, the mammal is ahuman.

In certain embodiments, the present invention provides a method oftreating a cancer or another proliferative disease, disorder orcondition, comprising administering a compound or composition of thepresent invention to a patient with a cancer or another proliferativedisease, disorder or condition. In certain embodiments, the method oftreating a cancer or other proliferative disorder comprisesadministering compounds and compositions of the present invention to amammal. In certain embodiments, the mammal is a human.

As used herein, the terms “treating a cancer” refers to the inhibitionof the growth, division, maturation or viability of cancer cells, and/orcausing the death of cancer cells, individually or in aggregate withother cancer cells, by cytotoxicity, nutrient depletion, or theinduction of apoptosis.

Examples of tissues containing cancerous cells whose proliferation isinhibited by the compounds and compositions described herein and againstwhich the methods described herein are useful include but are notlimited to breast, prostate, brain, blood, bone marrow, bone, liver,pancreas, skin, kidney, colon, ovary, lung, testicle, penis, thyroid,parathyroid, pituitary, thymus, retina, uvea, conjunctiva, spleen, head,neck, trachea, gall bladder, rectum, salivary gland, adrenal gland,throat, esophagus, lymph nodes, sweat glands, sebaceous glands, muscle,heart, and stomach.

In some embodiments, the cancer treated by compounds or compositions ofthe invention is a skin cancer, lung cancer, breast cancer, prostatecancer, leukemia, kidney cancer, esophageal cancer, brain cancer, bonecancer or colon cancer. In some embodiments, the cancer treated by thecompounds or compositions of the invention is an osteosarcoma, amelanoma or a prostate cancer.

In certain embodiments, the present invention provides a method oftreating a neurological disease, disorder or condition, comprisingadministering a compound or composition of the present invention to apatient with a neurological disease, disorder or condition. In certainembodiments, the method of treating a neurological disease, disorder orcondition comprises administering compounds and compositions of thepresent invention to a mammal. In certain embodiments, the mammal is ahuman. In certain embodiments, the neurological disease, disorder orcondition is pain or stroke.

In certain embodiments, the present invention provides a method oftreating an inflammatory disease, disorder or condition, comprisingadministering a compound or composition of the present invention to apatient with an inflammatory disease, disorder or condition. In certainembodiments, the method of treating an inflammatory disease, disorder orcondition comprises administering compounds and compositions of thepresent invention to a mammal. In certain embodiments, the mammal is ahuman. In certain embodiments, the neurological disease, disorder orcondition is asthma or rheumatoid arthritis.

In certain embodiments, the present invention provides a method oftreating a metabolic disease, disorder or condition, comprisingadministering a compound or composition of the present invention to apatient with a metabolic disease, disorder or condition. In certainembodiments, the method of treating a metabolic disease, disorder orcondition comprises administering compounds and compositions of thepresent invention to a mammal. In certain embodiments, the mammal is ahuman. In certain embodiments, the metabolic disease, disorder orcondition is diabetes. In some embodiments, the diabetes is type IIdiabetes.

In certain embodiments, the present invention provides a method oftreating opioid tolerance or dependence, comprising administering acompound or composition of the present invention to an opioid tolerantor dependent patient. In certain embodiments, the method of treatingopioid tolerance or dependence comprises administering compounds andcompositions of the present invention to a human. In some embodimentsthe opioid tolerance or dependence is morphine tolerance or dependence.

In certain embodiments, the present invention provides a method oftreating an ocular disease, disorder or condition, comprisingadministering a compound or composition of the present invention to apatient with an ocular disease, disorder or condition. In certainembodiments, the method of treating an ocular disease, disorder orcondition comprises administering compounds and compositions of thepresent invention to a mammal. In certain embodiments, the mammal is ahuman. In certain embodiments, the ocular disease, disorder or conditionis macular degeneration.

Depending upon the particular disease, disorder or condition to betreated, additional therapeutic agents, which are normally administeredto treat that condition, may be administered in combination withcompounds and compositions of this invention. As used herein, additionaltherapeutic agents that are normally administered to treat a particulardisease, or condition, are known as “appropriate for the disease, orcondition, being treated”.

In certain embodiments, a provided compound, or composition thereof, isadministered in combination with another inhibitor of CaMKII. In someembodiments, a provided compound, or composition thereof, isadministered in combination with one or more other therapeutic agents.Such CaMKII inhibitors include, but are not limited to CaM Kinase IICalmodulin Antagonist peptide, KN-93, and lavendustin C.

In certain embodiments, a provided compound, or a composition thereof,is administered in combination with another anti-cancer, cytotoxin, orchemotherapeutic agent.

In certain embodiments, the anti-cancer or chemotherapeutic agents usedin combination with compounds or compositions of the invention include,but are not limited to imatinib, nilotinib, gefitinib, sunitinib,carfilzomib, salinosporamide A, retinoic acid, cisplatin, carboplatin,oxaliplatin, mechlorethamine, cyclophosphamide, chlorambucil,ifosfamide, azathioprine, mercaptopurine, doxifluridine, fluorouracil,gemcitabine, methotrexate, tioguanine, vincristine, vinblastine,vinorelbine, vindesine, podophyllotoxin, etoposide, teniposide,tafluposide, paclitaxel, docetaxel, irinotecan, topotecan, amsacrine,actinomycin, doxorubicin, daunorubicin, valrubicin, idarubicin,epirubicin, plicamycin, mitomycin, mitoxantrone, melphalan, busulfan,capecitabine, pemetrexed, epothilones, 13-cis-Retinoic Acid, 2-CdA,2-Chlorodeoxyadenosine, 5-Azacitidine, 5-Fluorouracil, 5-FU,6-Mercaptopurine, 6-MP, 6-TG, 6-Thioguanine, Abraxane, Accutane®,Actinomycin-D, Adriamycin®, Adrucil®, Afinitor®, Agrylin®, Ala-Cort®,Aldesleukin, Alemtuzumab, ALIMTA, Alitretinoin, Alkaban-AQ®, Alkeran®,All-transretinoic Acid, Alpha Interferon, Altretamine, Amethopterin,Amifostine, Aminoglutethimide, Anagrelide, Anandron®, Anastrozole,Arabinosylcytosine, Ara-C, Aranesp®, Aredia®, Arimidex®, Aromasin®,Arranon®, Arsenic Trioxide, Arzerra™, Asparaginase, ATRA, Avastin®,Azacitidine, BCG, BCNU, Bendamustine, Bevacizumab, Bexarotene, BEXXAR®,Bicalutamide, BiCNU, Blenoxane®, Bleomycin, Bortezomib, Busulfan,Busulfex®, C225, Calcium Leucovorin, Campath®, Camptosar®,Camptothecin-11, Capecitabine, Carac™, Carboplatin, Carmustine,Carmustine Wafer, Casodex®, CC-5013, CCI-779, CCNU, CDDP, CeeNU,Cerubidine®, Cetuximab, Chlorambucil, Citrovorum Factor, Cladribine,Cortisone, Cosmegen®, CPT-11, Cytadren®, Cytosar-U®, Cytoxan®,Dacarbazine, Dacogen, Dactinomycin, Darbepoetin Alfa, Dasatinib,Daunomycin, Daunorubicin Hydrochloride, Daunorubicin Liposomal,DaunoXome®, Decadron, Decitabine, Delta-Cortef®, Deltasone®, Denileukin,Diftitox, DepoCyt™, Dexamethasone, Dexamethasone Acetate, DexamethasoneSodium Phosphate, Dexasone, Dexrazoxane, DHAD, DIC, Diodex, Docetaxel,Doxil®, Doxorubicin, Doxorubicin Liposomal, Droxia™, DTIC, DTIC-Dome®,Duralone®, Efudex®, Eligard™, Ellence™, Eloxatin™, Elspar®, Emcyt®,Epirubicin, Epoetin Alfa, Erbitux, Erlotinib, Erwinia L-asparaginase,Estramustine, Ethyol, Etopophos®, Etoposide, Etoposide Phosphate,Eulexin®, Everolimus, Evista®, Exemestane, Fareston®, Faslodex®,Femara®, Filgrastim, Floxuridine, Fludara®, Fludarabine, Fluoroplex®,Fluorouracil, Fluorouracil (cream), Fluoxymesterone, Flutamide, FolinicAcid, FUDR®, Fulvestrant, G-CSF, Gefitinib, Gemcitabine, Gemtuzumab,ozogamicin-Gemzar Gleevec™, Gliadel® Wafer, GM-CSF, Goserelin,Granulocyte-Colony Stimulating Factor, Granulocyte Macrophage ColonyStimulating Factor, Halotestin®, Herceptin®, Hexadrol, Hexalen®,Hexamethylmelamine, HMM, Hycamtin®, Hydrea®, Hydrocort Acetate®,Hydrocortisone, Hydrocortisone Sodium Phosphate, Hydrocortisone SodiumSuccinate, Hydrocortone Phosphate, Hydroxyurea, Ibritumomab,Ibritumomab, Tiuxetan, Idamycin®, Idarubicin Ifex®, IFN-alpha,Ifosfamide, IL-11, IL-2, Imatinib mesylate, Imidazole Carboxamide,Interferon alfa, Interferon Alfa-2b (PEG Conjugate), Interleukin-2,Interleukin-11, Intron A® (interferon alfa-2b), Iressa®, Irinotecan,Isotretinoin, Ixabepilone, Ixempra™, Kidrolase®, Lanacort®, Lapatinib,L-asparaginase, LCR, Lenalidomide, Letrozole, Leucovorin, Leukeran,Leukine™, Leuprolide, Leurocristine, Leustatin™, Liposomal Ara-C, LiquidPred®, Lomustine, L-PAM, L-Sarcolysin, Lupron®, Lupron Depot®,Matulane®, Maxidex, Mechlorethamine, Mechlorethamine Hydrochloride,Medralone®, Medrol®, Megace®, Megestrol, Megestrol Acetate, Melphalan,Mercaptopurine, Mesna, Mesnex™, Methotrexate, Methotrexate Sodium,Methylprednisolone, Meticorten®, Mitomycin, Mitomycin-C, Mitoxantrone,M-Prednisol®, MTC, MTX, Mustargen®, Mustine, Mutamycin®, Myleran®,Mylocel™, Mylotarg®, Navelbine®, Nelarabine, Neosar®, Neulasta™,Neumega®, Neupogen®, Nexavar®, Nilandron®, Nilotinib, Nilutamide,Nipent®, Nitrogen Mustard, Novaldex®, Novantrone®, Nplate, Octreotide,Octreotide acetate, Ofatumumab, Oncospar®, Oncovin®, Ontak®, Onxal™,Oprelvekin, Orapred®, Orasone®, Oxaliplatin, Paclitaxel, PaclitaxelProtein-bound, Pamidronate, Panitumumab, Panretin®, Paraplatin®,Pazopanib, Pediapred®, PEG Interferon, Pegaspargase, Pegfilgrastim,PEG-INTRON™, PEG-L-asparaginase, PEMETREXED, Pentostatin, PhenylalanineMustard, Platinol®, Platinol-AQ®, Prednisolone, Prednisone, Prelone®,Procarbazine, PROCRIT®, Proleukin®, Prolifeprospan 20 with CarmustineImplant, Purinethol®, Raloxifene, Revlimid®, Rheumatrex®, Rituxan®,Rituximab, Roferon-A® (Interferon Alfa-2a), Romiplostim, Rubex®,Rubidomycin hydrochloride, Sandostatin®, Sandostatin LAR®, Sargramostim,Solu-Cortef®, Solu-Medrol®, Sorafenib, SPRYCEL™, STI-571, Streptozocin,SU11248, Sunitinib, Sutent®, Tamoxifen, Tarceva®, Targretin®, Tasigna®,Taxol®, Taxotere®, Temodar®, Temozolomide, Temsirolimus, Teniposide,TESPA, Thalidomide, Thalomid®, TheraCys®, Thioguanine, ThioguanineTabloid®, Thiophosphoamide, Thioplex®, Thiotepa, TICE®, Toposar®,Topotecan, Toremifene, Torisel®, Tositumomab, Trastuzumab, Treanda®,Tretinoin, Trexall™, Trisenox®, TSPA, TYKERB®, VCR, Vectibix™, Velban®,Velcade®, VePesid®, Vesanoid®, Viadur™, Vidaza®, Vinblastine,Vinblastine Sulfate, Vincasar Pfs®, Vincristine, Vinorelbine,Vinorelbine tartrate, VLB, VM-26, Vorinostat, Votrient, VP-16, Vumon®,Xeloda®, Zanosar®, Zevalin™, Zinecard®, Zoladex®, Zoledronic acid,Zolinza, Zometa®, or combinations of any of the above.

In certain embodiments, a combination of 2 or more therapeutic agentsmay be administered together with compounds of the invention. In certainembodiments, a combination of 3 or more therapeutic agents may beadministered with compounds of the invention.

Other examples of agents the inhibitors of this invention may also becombined with include, without limitation: vitamins and nutritionalsupplements, cancer vaccines, treatments for neutropenia (e.g. G-CSF,filgrastim, lenograstim), treatments for thrombocytopenia (e.g. bloodtransfusion, erythropoietin), antiemetics (e.g. 5-HT₃ receptorantagonists, dopamine antagonists, NK1 receptor antagonists, histaminereceptor antagonists, cannabinoids, benzodiazepines, oranticholinergics), treatments for Alzheimer's Disease such as Aricept®and Excelon®; treatments for Parkinson's Disease such asL-DOPA/carbidopa, entacapone, ropinrole, pramipexole, bromocriptine,pergolide, trihexephendyl, and amantadine; agents for treating MultipleSclerosis (MS) such as beta interferon (e.g., Avonex® and Rebif®),Copaxone®, and mitoxantrone; treatments for asthma such as albuterol andSingulair®; agents for treating schizophrenia such as zyprexa,risperdal, seroquel, and haloperidol; anti-inflammatory agents such ascorticosteroids, TNF blockers, IL-1 RA, azathioprine, cyclophosphamide,and sulfasalazine; immunomodulatory and immunosuppressive agents such ascyclosporin, tacrolimus, rapamycin, mycophenolate mofetil, interferons,corticosteroids, cyclophophamide, azathioprine, and sulfasalazine;neurotrophic factors such as acetylcholinesterase inhibitors, MAOinhibitors, interferons, anti-convulsants, ion channel blockers,riluzole, and anti-Parkinsonian agents; agents for treatingcardiovascular disease such as beta-blockers, ACE inhibitors, diuretics,nitrates, calcium channel blockers, and statins, fibrates, cholesterolabsorption inhibitors, bile acid sequestrants, and niacin; agents fortreating liver disease such as corticosteroids, cholestyramine,interferons, and anti-viral agents; agents for treating blood disorderssuch as corticosteroids, anti-leukemic agents, and growth factors;agents for treating immunodeficiency disorders such as gamma globulin;and anti-diabetic agents such as biguanides (metformin, phenformin,buformin), thiazolidinediones (rosiglitazone, pioglitazone,troglitazone), sulfonylureas (tolbutamide, acetohexamide, tolazamide,chlorpropamide, glipizide, glyburide, glimepiride, gliclazide),meglitinides (repaglinide, nateglinide), alpha-glucosidase inhibitors(miglitol, acarbose), incretin mimetics (exenatide, liraglutide,taspoglutide), gastric inhibitory peptide analogs, DPP-4 inhibitors(vildagliptin, sitagliptin, saxagliptin, linagliptin, alogliptin),amylin analogs (pramlintide), and insulin and insulin analogs.

In certain embodiments, compounds of the present invention, or apharmaceutically acceptable composition thereof, are administered incombination with antisense agents, a monoclonal or polyclonal antibodyor an siRNA therapeutic.

Those additional agents may be administered separately from an inventivecompound-containing composition, as part of a multiple dosage regimen.Alternatively or in addition to those additional agents administeredseparately, those agents may be part of a single dosage form, mixedtogether with a compound of this invention in a single composition. Ifadministered as part of a multiple dosage regime, the two active agentsmay be submitted simultaneously, sequentially or within a period of timefrom one another, normally within five hours from one another.

As used herein, the term “combination,” “combined,” and related termsrefers to the simultaneous or sequential administration of therapeuticagents in accordance with this invention. For example, a compound of thepresent invention may be administered with another therapeutic agentsimultaneously or sequentially in separate unit dosage forms or togetherin a single unit dosage form. Accordingly, the present inventionprovides a single unit dosage form comprising a compound of formula I,I-a, II, III, IV, V, VI, VII, VIII, or IX, an additional therapeuticagent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

The amount of both, an inventive compound and additional therapeuticagent (in those compositions which comprise an additional therapeuticagent as described above) that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. Preferably,compositions of this invention should be formulated so that a dosage ofbetween 0.01-100 mg/kg body weight/day of an inventive can beadministered.

In those compositions which comprise an additional therapeutic agent,that additional therapeutic agent and the compound of this invention mayact synergistically. Therefore, the amount of additional therapeuticagent in such compositions will be less than that required in amonotherapy utilizing only that therapeutic agent. In such compositionsa dosage of between 0.01-100 μg/kg body weight/day of the additionaltherapeutic agent can be administered.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered in a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

EXEMPLIFICATION

As depicted in the Examples below, in certain exemplary embodiments,compounds are prepared according to the following general procedures. Itwill be appreciated that, although the general methods depict thesynthesis of certain compounds of the present invention, the followinggeneral methods, and other methods known to one of ordinary skill in theart, can be applied to all compounds and subclasses and species of eachof these compounds, as described herein.

In certain embodiments, compounds of formula I-a are prepared accordingto the procedure outlined in Scheme 1.

Depending on the nature the of the functional groups on specificinstances of R^(2′) and R^(3′), functional groups on those substituentswere modified by protecting groups according to standard proceduresknown in the art prior to the Suzuki coupling step, and an optionaldeprotection step was used to remove those protecting groups subsequentto the Suzuki coupling. In some embodiments, primary amine functionalgroups on substituents R^(2′) and R^(3′) may be protected, prior to thecoupling step, thereby forming modified groups R^(2′)-P and R^(3′)-Prespectively, wherein P is a protecting group suitable for primaryamines. In some embodiments, protecting group P is a carbamate (e.g.Cbz, BOC, methylcarbamate, ethylcarbamate). In some embodiments,protecting group P is a BOC (t-butoxycarbonyl) group. Methods for theintroduction and removal of such primary amine protecting groups arewell known in the art, as exemplified by “Greene's Protective Groups inOrganic Synthesis” Wuts P G M, Greene, T W, 4^(th) Ed., 2006,Wiley-Interscience, the entirety of which is incorporated herein byreference. In certain embodiments, compounds and synthetic intermediatesthereof of the present invention may be prepared according to methodsknown to one of ordinary skill in the art. One of ordinary art willappreciate that compounds of formula I bearing additional substituentsR^(4′), R^(5′), R^(6′), R⁵, and R⁸, may be prepared according to similarprocedures to those described above for formula I-a.

The following examples illustrate methods utilized for the preparationof structures relevant to the present invention. Throughout theseexamples, certain equipment, HPLC columns and solvent systems wereutilized in the execution of reactions and the purification of reactionproducts. Accordingly, microwave reactions were carried out utilizing anAntonPar, Monowave 300 microwave reactor. Preparative HPLC purificationswere carried out utilizing a Shimadzu [Prominence LC-20AP], equippedwith a Discovery C-18 column (50×21.2 mm, 5μ) utilizing the followingmethod: Solvent A=Acetonitrile, Solvent B=Water; Gradient=95% solvent Bto 10% solvent B over 20 min with a flow rate of 10 mL/min. AnalyticalLCMS data were acquired using a Shimadzu [LCMS-2020] equipped with aSHIMPAK, XR ODS-II column (50×2 mm) utilizing the following method: FlowRate=0.2 mL/min, Solvent A=Acetonitrile, Solvent B=0.1% TFA in water;Gradient=Initial 95% of solvent B to 10% solvent B over 10 min followedby 10% solvent B for an additional 10 min.

Example 1

An exemplary procedure for the preparation of compound 9 (I-7) follows.

Synthesis of 1-(pyridin-2-yl)-1H-benzo[d][1,2,3]triazole (3)

A suspension of 1H-benzo-[1,2,3]triazole (1, 40 g, 335 mmol) and2-bromopyridine (2, 105 g, 671 mmol) in toluene (160 mL) was heated atreflux for 18 h after which, the reaction mixture was poured into EtOAc(1 L). The resulting white solid precipitate was dissolved by additionof aqueous KOH (10%, 85 mL). The phases were separated, and the organiclayer was washed with aqueous KOH (10%, 2×250 mL). The organic layer wasdried over anhydrous Na₂SO₄, filtered and concentrated to dryness. Theisolated solids were recrystallized from CH₃OH giving a 62% yield ofcompound 3. ¹H NMR (500 MHz, CDCl₃) δ ppm: 8.65 (d, 1H), 8.62 (d, 1H),8.31 (d, 1H), 8.11 (d, 1H), 7.94 (m, 1H), 7.59 (t, 1H), 7.46 (t, 1H),7.33 (m, 1H); Mass (m/z): 197.2 (M+H).

Synthesis of 9H-pyrido[2,3-b]indole (4)

To compound 3 (40 g, 203 mmol) was added polyphosphoric acid (160 g)pre-heated to 160° C. Once gas evolution was complete, H₂O (900 mL) wasadded and the pH of the solution was adjusted >10 by addition of aqueousNaOH (10M). The mixture was then sonicated at 50° C. until the reactionmass was fully suspended. The suspension was poured into H₂O (500 mL)and cooled to room temperature. After 20 min, the resulting solid wascollected by filtration, washed with H₂O (2×150 ML) and dried undervacuum giving a 35% yield of crude compound 4. ¹H NMR (500 MHz, DMSO-d₆)δ ppm: 11.72 (br, 1H), 8.44 (d, 1H), 8.37 (d, 1H), 8.11 (d, 1H), 7.47(dd, 1H), 7.38-7.44 (m, 1H), 7.16-7.20 (m, 1H), 7.15 (d, 1H); Mass(m/z): 169.2 (M+H)

Synthesis of 9H-pyrido[2,3-b]indole 1-oxide (5)

Aqueous H₂O₂ (35%, 2.8 g, 83 mmol) was added dropwise to a solution ofcrude compound 4 (2 g, 11.9 mmol) in CH₃COOH (17 mL). The reactionmixture was refluxed for 4 h. Additional aqueous H₂O₂ (35%, 1 mL) wasadded dropwise and refluxing was continued for an additional 2 h. Thesolvent was then removed under vacuum and pH of the oily residue wasadjusted to 8 on treatment with saturated aqueous K₂CO₃. The resultingsolution was stirred overnight. The resulting solid was collected byfiltration, washed with H₂O and dried under giving a 67% yield ofcompound 5. ¹H NMR (500 MHz, DMSO-d₆) δ ppm: 12.58 (br, 1H), 8.34(d,1H), 8.20 (dd, 2H), 7.56 (m, 2H), 7.30 (t, 1H), 7.23 (t, 1H). Mass(m/z): 185.2 (M+H).

Synthesis of 4-bromo-9H-pyrido[2,3-b]indole (6)

A suspension of compound 5 (1 g, 5.5 mmol) in anhydrous DMF (10 mL) wascooled to 0° C. and POBr₃ (3.66 g, 12.8 mmol) was added dropwise withstirring. The reaction was stirred for 24 h at room temperature and thenpoured into H₂O (20 mL). After cooling to 0-5° C. the pH of the solutionwas adjusted to 8-10 with aqueous KOH (10%). After stirring for anadditional 15 min, the resulting precipitate was collected byfiltration, washed with H₂O 15 mL and dried under vacuum. The crudematerial was purified on silica gel (20% EtOAc in Hexane) giving a 50%yield of compound 6. ¹H NMR (500 MHz, DMSO-d₆) δ ppm: 12.19 (br, 1H),8.46(d, 1H), 8.27 (d, 1H), 7.57 (m, 2H), 7.46 (d, 1H), 7.32 (m, 1H).Mass (m/z): 247.2 (M+H).

Synthesis of tert-butyl2-(3-(9H-pyrido[2,3-b]indol-4-yl)phenylamino)ethylcarbamate (8)

A solution of compound 6 (250 mg, 1.01 mmol), boronate ester 7 (550 mg,1.52 mmol), aqueous sodium carbonate (2.0M, 1 mL, 2.0 mmol) and dioxane(4 mL) was purged with nitrogen and Pd(PPh₃)₄ (58 mg, 0.05 mmol) wasadded. The resulting mixture was stirred in a microwave reactor at 140°C. for 45 min. After cooling to room temperature, the mixture was pouredinto water (30 mL) and washed with EtOAc (2×50 mL). The combined organicphases were dried over anhydrous Na₂SO₄, filtered, and concentrated invacuo. The crude product was purified on silica gel (40% EtOAc inhexane) giving a 40% yield of compound 8. Mass (m/z): 403.4 (M+H).

Synthesis of N1-(3-(9H-pyrido[2,3-b]indol-4-yl)phenyl)ethane-1,2-diamine(9, Compound I-7)

Trifluoroacetic acid (114 mg, 1.0 mmol) was added dropwise to asuspension of compound 8 (100 mg, 0.2 mmol) in anhydrous dichloromethane(3 mL). After stirring for 2 h at room temperature, the mixture wasconcentrated in vacuo giving 110 mg of compound 9. This material wasstirred with desalting resin (MP-carbonate, 150 mg) in MeOH (4 mL) for 2h. The resin was removed by filtration and the filtrate was concentratedto dryness. Purification of the residue by preparative HPLC gave 22 mg(30% yield) of pure compound 9. ¹H NMR (500 MHz, DMSO-d₆) δ ppm: 11.9(br, 1H), 8.41(d, 1H), 7.8 (br, 2H), 7.64 (d, 1H), 7.49 (d, 1H), 7.39(t, 1H), 7.33 (m, 1H), 7.04 (m, 2H), 6.78-6.85(m, 3H), 6.0 (br, 1H),3.25(t, 2H), 2.97(t, 2H). Mass (m/z): 303.2 (M+H). Purity: 98.4% byHPLC.

Synthesis of tert-butyl2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylamino)-ethylcarbamate(7)

Synthesis of tert-butyl 2-(3-bromophenylamino)ethylcarbamate (12)

A mixture of 1,3-dibromobenzene 10 (2.2 g, 9.36 mmol), tert-butyl2-aminoethylcarbamate 11 (1 g, 6.2 mmol), CuI (237 mg, 1.2 mmol),L-proline (215 mg, 1.8 mmol) and potassium carbonate (1.7 g, 12.4 mmol)in DMSO (20 mL) was placed under N₂ and heated to 90° C. for 14 h. Thereaction mixture was cooled to room temperature and concentrated underreduced pressure to dryness. The resulting residue was dissolved inethyl acetate (100 mL), washed with water (2×25 mL) and washed withbrine (2×25 mL). After drying over anhydrous Na₂SO₄, the organic phasewas filtered and concentrated to dryness. The residue was purified onsilica gel (30% EtOAc/hexane) giving compound 12 (1 g, 50% yield) as abrown solid. ¹H NMR (500 MHz, CDCl₃) δ ppm: 7.00 (t, 1H), 6.81(d, 1H),6.72 (s, 1H), 6.51 (d, 1H), 4.75 (br, 1H), 4.2 (br, 1H), 3.35 (t, 2H),3.22 (t, 2H), 1.45 (s, 9H). Mass (m/z): 315.1and 317.1.

Synthesis of tert-butyl2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylamino)-ethylcarbamate(7)

A mixture of tert-butyl 2-(3-bromophenylamino)ethylcarbamate 12 (1 g,3.17 mmol), bis(pinacolato)diboron 13 (1.2 g, 4.76 mmol),palladium(II)acetate (50 mg, 0.2 mmol), BINAP (100 mg, 0.15 mmol) andcesium carbonate (2 g, 6.3 mmol) in dioxane (20 mL) was stirred at 90°C. under N₂ for 14 h. After cooling to room temperature, the mixture wasfiltered through Celite and washed with EtOAc. The filtrate wasconcentrated to dryness giving crude compound 7 (1.2 g) which was usedwithout purification. Mass (m/z): 363.3 (M+H).

Example 2 Synthesis ofN1-(3-(9H-pyrido[2,3-b]indol-4-yl)phenyl)propane-1,3-diamine (9a,Compound I-8)

Compounds 8a and 9a were prepared from compound 6 and compound 7aaccording to the protocols described for the preparation of compound 8and compound 9.

Synthesis of tert-butyl2-(3-(9H-pyrido[2,3-b]indol-4-yl)phenylamino)propylcarbamate (8a)

Compound 8a was prepared according to the procedure for compound 8(Example 1) using compound 6 (100 mg, 0.405 mmol), compound 7a (228 mg,0.60 mmol), Pd(PPh₃)₄ (23 mg, 0.025 mmol) and proportionate molarequivalents of aqueous sodium carbonate and dioxane. 80 mg (47% yield)of compound 8a were isolated. Mass (m/z): 417.4 (M+H). Purity: 98.2% byHPLC.

Synthesis ofN1-(3-(9H-pyrido[2,3-b]indol-4-yl)phenyl)propane-1,2-diamine (9a,Compound I-8)

Compound 9a was prepared according the procedure for compound 9(Example 1) using compound 8a (80 mg, 0.19 mmol) and trifluoroaceticacid (109 mg, 0.95 mmol). 12 mg (20% yield) of compound 9a wereisolated. ¹H NMR (500 MHz, DMSO-d₆) δ ppm: 11.91 (br, 1H), 8.41(d, 1H),7.7 (br, 2H), 7.64 (d, 1H), 7.49 (d, 1H), 7.41 (t, 1H), 7.31 (t, 1H),7.04-7.06 (m, 2H), 6.83(d, 1H), 6.78 (d, 1H), 5.94(br, 1H),3.31(t,2H),2.90(t,2H),1.83-1.85(m,2H). Mass (m/z): 317.3 (M+H).

Synthesis of tert-butyl3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylamino)-propylcarbamate(7a)

Compounds 12a and 7a were prepared from 1,3-dibromobenzene 10 and theappropriate mono-Boc protected diamine 11a according to the protocolsdescribed for the preparation of compound 12 and compound 7.

Conversion of Compound 10 to Compound 12a:

Compound 12a was prepared according to the procedure for compound 12(Example 1) using compound 11a (350 mg, 2.96 mmol) and proportionatemolar equivalents of compound 10, CuI, L-proline, potassium carbonateand DMSO. Compound 12a was isolated in 25% yield. ¹H NMR (500 MHz,CDCl₃) δ ppm: 6.99 (t, 1H), 6.77(d, 1H), 6.73 (s, 1H), 6.50 (d, 1H),3.22 (d, 2H),3.15 (t, 2H), 1.75 (m, 2H), 1.45 (s, 9H). Mass (m/z):329.1, 331.1.

Conversion of Compound 12a to Compound 7a:

Compound 7a was prepared according to the procedure for compound 7(Example 1) using 12a (1 g, 3.03 mmol), bis(pinacolato)diboron 13 (1.15g, 4.55 mmol), palladium(II)acetate (50 mg, 0.2 mmol), BINAP (100 mg,0.15 mmol) and cesium carbonate (1.98 g, 6.07 mmol) in dioxane (20 mL).crude compound 7a (1.25 g) was used without purification. Mass (m/z):377.3 (M+H).

Example 3 Synthesis of 3-(9H-pyrido[2,3-b]indol-4-yl)aniline (9b,Compound I-6)

Compounds 8b and 9b were prepared from 6 and compound 7b according tothe protocols described for the preparation of compound 8 and compound9.

Conversion of Compound 6 to Compound 8b

Compound 8b was prepared according to the procedure for compound 8(Example 1) using compound 6 (100 mg, 0.405 mmol), compound 7b (194 mg,0.60 mmol), Pd(PPh₃)₄ (23 mg, 0.025 mmol) and proportionate molarequivalents of aqueous sodium carbonate and dioxane. 67 mg (46% yield)of compound 8b were isolated. Mass (m/z): 360.4 (M+H). Purity: 97.12% byHPLC.

Conversion of Compound 8b to Compound 9b (I-6):

Compound 9b was prepared according to the procedure for compound 9(Example 1) using compound 8b (67 mg, 0.18 mmol) and trifluoroaceticacid (106 mg, 0.90 mmol). 22 mg (45% yield) of compound 9b wereisolated. ¹H NMR (500 MHz, DMSO-d⁶) δ ppm: 11.89 (br, 1H), 8.41(d, 1H),7.66 (d, 2H), 7.50 (d, 1H), 7.48 (d, 1H), 7.39 (t, 1H), 7.22 (t, 1H),7.03 (m, 2H), 6.83(s, 1H), 6.75 (m, 2H), 5.30(br, 2H). Mass (m/z): 260.3(M+H).

Synthesis of tert-butyl3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate (7b)

Synthesis of tert-butyl 3-bromophenylcarbamate (12b)

To a solution of 3-bromoaniline (300 mg, 1.7 mmole) in dichloromethane(5 mL) at 0° C. was added triethylamine (3.4 mL, 0.47 mmoles, 2.0 eq.).After stirring for 5-10 min, di-tert-butyl dicarbonate (452 mg, 2 moles,1.2 eq.) was added slowly and the reaction was allowed to stir at roomtemperature for 12 h. The reaction was then concentrated to dryness andthe residue was purified on silica gel (5% EtOAc/hexane) giving compound12b (272 mg, 60% Yield). ¹H NMR (500 MHz, CDCl₃) δ PPM: 7.65 (s,1H),7.25(d, 1H), 7.11-7.13 (m, 2H), 6.51 (br, 1H), 1.46 (s, 9H).

Conversion of Compound 12b to Compound 7b:

Compound 7b was prepared according to the procedure for compound 7(Example 1) using compound 12b (200 mg) and proportionate molarequivalents of compound 13, palladium(II)acetate, BINAP, cesiumcarbonate and dioxane. 300 mg of crude 7b were isolated and used withoutpurification.

Example 4 Synthesis of (3-(9H-pyrido[2,3-b]indol-4-yl)phenyl)methanamine(9c, Compound I-3)

Compounds 8c and 9c were prepared from compound 6 and compound 7caccording to the protocols described for the preparation of compound 8and compound 9.

Conversion of Compound 6 to Compound 8c:

Compound 8c was prepared according to the procedure for compound 8(Example 1) using compound 6 (100 mg, 0.405 mmol), compound 7c (199 mg,0.60 mmol), Pd(PPh₃)₄ (23 mg, 0.025 mmol) and proportionate molarequivalents of aqueous sodium carbonate and dioxane. 75 mg (50% yield)of compound 8c were isolated. Mass (m/z): 374.4 (M+H). Purity: 95.5% byHPLC.

Conversion of Compound 8c to Compound 9c (I-3):

Compound 9c was prepared according to the procedure for compound 9(Example 1) using compound 8c (75 mg, 0.2 mmol), and trifluoroaceticacid (118 mg, 0.90 mmol). 12 mg (20% yield) of compound 9a wereisolated. ¹H NMR (500 MHz, DMSO-d⁶) δ PPM: 11.89 (br, 1H), 8.48(d, 1H),7.77 (s, 1H), 7.65-7.68 (m, 3H), 7.53 (dd, 2H), 7.42 (t, 1H), 7.10 (d,1H), 7.09 (t, 1H),4.12(s,2H). Mass (m/z): 274.3 (M+H).

Synthesis of tert-butyl3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzylcarbamate (7c)

Compounds 12c and 7c were prepared from 3-bromobenzylamine hydrochlorideand compound 13 according to the protocols described for the preparationof compound 12b and compound 7b.

Synthesis of tert-butyl 3-bromobenzylcarbamate (12c)

Compound 12c was prepared according to the procedure for compound 12b(Example 3) using 3-bromobenzylamine hydrochloride (125 mg, 0.56 mmol)and proportionate molar equivalents of dichloromethane, triethylamineand di-tert-butyl dicarbonate. 98 mg (60% yield) of compound 12c wereisolated.

Conversion of 12c to 7c:

Compound 7c was prepared according to the procedure for compound 7(Example 1) using compound 12c (90 mg) and proportionate molarequivalents of compound 13, palladium(II)acetate, BINAP, cesiumcarbonate and dioxane. 199 mg of crude 7c were isolated and used withoutpurification.

Example 5 Synthesis of 1-(3-(9H-pyrido[2,3-b]indol-4-yl)phenyl)guanidine(14, Compound I-4)

To a solution of 9b (30 mg, 0.115 mmol) in ethanol (2 mL) was addedNH₂CN (29 mg, 6 eq.). The mixture was refluxed for 2 days after which,the volatiles were removed under reduced pressure. The resulting cruderesidue was dissolved in water and the pH was adjusted to ˜10 usingaqueous 2N NaOH. The aqueous mixture was extracted with EtOAc (5 mL).The organic phase was dried over Na₂SO₄, concentrated and purified bypreparative HPLC giving compound 14 (4 mg, 12% yield) as a yellow solid.Mass (m/z): 302.3(M+H). Purity: 90.05% by HPLC.

Example 6 Synthesis of 1-(3-(9H-pyrido[2,3-b]indol-4-yl)benzyl)guanidine(14a, Compound I-5)

Compound 14a was prepared according to the preparation of compound 14(Example 5) using compound 9c (40 mg, 0.146 mmol) and NH₂CN (35 mg, 6eq.). 4 mg (8.6% yield) of compound 14a were isolated. Mass (m/z): 316.4(M+H). Purity: 98.1% by HPLC.

Example 7 Synthesis of Compound 18 (I-2)

Preparation of Compound 16:

A Solution of compound 6 (250 mg, 1.01 mmol), 3-hydroxyphenylboronicacid 15 (167 mg, 1.2 mmol) and aqueous sodium carbonate (2 M, 1 mL, 2.0mmol) in dioxane (4 mL) was purged with nitrogen. Pd(PPh₃)₄ (58 mg, 0.05mmol) was added and the resulting mixture was stirred at 90° C. for 5 h.After cooling to room temperature, the mixture was poured into water (30mL) and extracted with EtOAc (2×50 mL). The combined organic phases weredried over Na₂SO₄, filtered, and concentrated to dryness. The residuewas purified on silica gel (40% EtOAc/hexane) giving compound 16 (50%yield). ¹H NMR (500 MHz, DMSO-d₆) δ ppm: 11.89 (br, 1H), 9.68(br, 1H),8.42 (d,1H), 7.58 (d, 1H), 7.49 (d, 1H), 7.39 (m, 2H), 7.04 (m, 4H),6.92 (d, 1H). Mass (m/z): 261.3 (M+H).

Synthesis of 3-(9H-pyrido[2,3-b]indol-4-yl)phenyltrifluoromethanesulfonate (17)

To a solution of compound 16 (200 mg, 0.7 mmole) in dichloromethane (5mL) at 0° C. was added triethylamine (0.26 mL, 1.9 mmole). Afterstirring for 5-10 min, trifilic anhydride (0.2 mL, 1.5 mmole) was addeddrop-wise. Stirring was continued at room temperature for 2 h afterwhich, the reaction was concentrated to dryness and the residue waspurified on silica gel (20% EtOAc/hexane) giving compound 17 (150 mg,33% Yield). Mass (m/z): 393.3 (M+H).

Synthesis of 4-(3-(1H-imidazol-1-yl)phenyl)-9H-pyrido[2,3-b]indole (18,I-2)

A mixture of compound 17 (150 mg, 0.38 mmol), imadazole (77 mg, 1.14mmol), palladium(II)acetate (9 mg, 10 mol %), BINAP (24 mg, 10 mol %)and cesium carbonate (250 mg, 0.76 mmol) in THF (4 mL) was stirred in amicrowave reactor at 150° C. for 45 min. The mixture was filteredthrough Celite and the Celite pad was washed with EtOAc. The combinedfiltrate was concentrated to dryness and the crude product was purifiedby preparative HPLC to give compound 18 (7 mg, 5.8% yield) as an offwhite solid. Mass (m/z): 311.2 (M+H). Purity: 99.2% by HPLC.

Example 8 Synthesis of Compound 23 (I-1)

Synthesis of tert-butyl 4-(3-bromophenyl)piperazine-1-carboxylate (20)

A mixture of 1,3-dibromobenzene 10 (1.5 g, 6.35 mmol), tert-butylpiperazine-1-carboxylate 19 (1.18 g, 6.35 mmol), palladium(II)acetate(14 mg, 0.06 mmol), BINAP (79 mg, 0.12 mmol) and cesium carbonate (4g,12.7 mmol) in dioxane (25 mL) was stirred at 90° C. under N₂ for 16 h.The mixture was filtered through Celite and the Celite pad was washedwith EtOAc. The combined filtrate was concentrated to dryness and thecrude product was purified on silica gel (5% EtoAc/hexane) givingcompound 20 (500 mg, 23% yield) as an off white solid. ¹H NMR (500 MHz,CDCl₃) δ ppm: 7.10 (t, 1H), 7.08 (s, 1H), 6.98 (d,1H), 6.81 (d, 1H),3.55 (t,4H), 3.13 (t,4H), 1.45 (s,9H).

Synthesis of 3-(4-(tert-butoxycarbonyl)piperazin-1-yl)phenylboronic acid(21)

To a solution of compound 20 (500 mg, 1.46 mmol) in anhydrous THF (5 mL)at −78° C. was added n-BuLi (2.5M in THF, 0.05 mL, 2.19 mmol) dropwise.After stirring at −78° C. for 45 min, triisopropylborate (0.68 mL, 2.93mmol) was added dropwise. Stirring was continued for 2 h at −78° C. andthe reaction was allowed to warm to room temperature overnight. Thereaction was quenched with aqueous saturated NH₄Cl (5 mL) and extractedwith EtOAc (2×25 mL). The combined organic phases were dried overNa₂SO₄, filtered and concentrated to dryness. The residue was purifiedon silica gel (5% MeOH/dichloromethane) giving compound 21 (150 mg, 33%yield). ¹H NMR (500 MHz, CDCl₃) δ ppm: 7.10 (t, 1H), 7.08 (s, 1H), 6.98(d,1H), 6.81(d, 1H), 3.55 (t,4H), 3.13 (t,4H), 1.45 (s,9H). Mass (m/z):307.3 (M+H).

Synthesis of tert-butyl4-(3-(9H-pyrido[2,3-b]indol-4-yl)phenyl)piperazine-1-carboxylate (22)

Compound 22 was prepared according to the procedure for compound 16(Example 7) using compound 6 (100 mg, 0.405 mmol), compound 21 (148 mg,0.48 mmol) and Pd(PPh₃)₄ (23 mg, 0.025 mmol). 90 mg (52% yield) ofcompound 22 were isolated. ¹H NMR (500 MHz, DMSO-d₆) δ ppm: 11.94 (br,1H), 8.45 (d,1H), 7.50-7.58(m, 3H), 7.42 (t, 1H), 7.25 (s, 1H),7.19-7.21 (m, 3H), 7.03 (t, 1H),3.20(t,4H),3.47(t,4H).1.41(s,9H). Mass(m/z): 429.3 (M+H).

Synthesis of 4-(3-(piperazin-1-yl)phenyl)-9H-pyrido[2,3-b]indole (23,I-1)

Compound 23 was prepared according to the procedure for compound 9(Example 1) using compound 22 (75 mg, 0.2 mmol) and trifluoroacetic acid(118 mg, 0.90 mmol). 12 mg (20% yield) of compound 23 were isolated. ¹HNMR (500 MHz, DMSO-d⁶) δ ppm: 11.97 (br, 1H),8.87(b, 1H), 8.45 (d,1H),7.54-7.58(m, 3H), 7.42 (d, 1H), 7.25 (s, 1H), 7.19-7.21 (m, 3H), 7.03(t, 1H),3.26(t,4H),3.37(t,4H). Mass (m/z):

Example 9 Synthesis of Compound 28 (I-9)

Synthesis of tert-butyl 2-(3-bromophenoxy)ethylcarbamate (25)

A mixture of 3-bromophenol 24 (200 mg, 1.15 mmol), tert-butyl2-bromoethylcarbamate (257 mg, 1.15 mmol) and cesium carbonate (753 mg,2.3 mmol) in DMF (5 mL) was stirred at 90° C. under N₂ for 5 h. Themixture was filtered through Celite and the Celite pad was washed withDMF (3 mL). The filtrate was concentrated to dryness and the crudeproduct was purified on silica gel (20% EtOAc/hexane) giving compound 25in 60% yield. ¹H NMR (500 MHz, CDCl₃) δ PPM: 7.05-7.19 (m, 3H), 6.80(dd, 1H), 5.0 (br, 1H), 3.98 (t, 2H), 3.52 (t, 2H), 1.46 (s, 9H).

Synthesis of tert-butyl2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethylcarbamate(26)

A mixture of tert-butyl 2-(3-bromophenoxy)ethylcarbamate 25 (300 mg,0.94 mmol), bis(Pinacolato)diboron (360 mg, 1.42 mmol),Palladium(II)acetate (12 mg, 0.05 mmol), BINAP (22 mg, 0.03 mmol) andCesium carbonate (590 mg,1.8 mmol) in dioxane (20 mL) was stirred at 90°C. under N₂ for 16 h. The mixture was filtered through Celite and theCelite pad was washed with EtOAc. The filtrate was concentrated todryness giving crude compound 26 which was used in the next step withoutfurther purification.

Synthesis of tert-butyl 2-(3-(9H-pyrido[2,3-b]indol-4-yl)phenoxy)ethylcarbamate (27)

A mixture of compound 6 (100 mg, 0.40 mmol), compound 26 (220 mg, 0.60mmol), aqueous sodium carbonate (2 M, 1 mL, 2.0 mmol) in dioxane (4 mL)was purged with nitrogen. Pd(PPh₃)₄ (23 mg, 0.02 mmol) was added and theresulting mixture was stirred in a microwave reactor at 140° C. for 45min. After cooling to room temperature, the mixture was poured intowater (10 mL) and extracted with EtOAc (2×25 mL). The combined organiclayers were dried over Na₂SO₄, filtered, and concentrated to dryness.The residue was purified on silica gel (40% EtOAc/hexane) givingcompound 27 (50% yield). Mass (m/z): 404.4 (M+H).

Synthesis of 2-(3-(9H-pyrido[2,3-b]indol-4-yl)phenoxy)ethanamine (28,Compound I-9)

To a solution of compound 27 (100 mg, 0.24 mmol) in anhydrousdichloromethane (3 mL) was added trifluoroacetic acid (141 mg, 1.2 mmol)dropwise. After stirring at room temperature for 2 h, the reaction wasconcentrated to dryness. The residue was stirred with MP-carbonate resin(150 mg) in MeOH (5 mL) for 2 h. The resin was removed by filtration.The filtrate was concentrated to dryness and the residue was purified bypreparative HPLC giving compound 28 (14 mg, 18% yield) as a off whitesolid. ¹H NMR (500 MHz, DMSO-d₆) δ PPM: 11.98 (br, 1H), 8.43(d, 1H),7.98 (br, 2H), 7.55 (m, 3H), 7.40 (t, 1H), 7.29 (d, 1H), 7.24 (s, 1H),7.18 (d, 2H), 7.08(d, 3H), 7.02 (t, 1H). Mass (m/z): 304.3 (M+H).

Example 10 Synthesis of Compound 28a (I-10)

Compound 28a was prepared from compound 24 according to protocolsdescribed for the preparation of compounds 25, 26, 27 and 28.

Synthesis of tert-butyl 3-(3-bromophenoxy)propylcarbamate (25a)

Compound 25a was prepared according to the procedure for compound 25(Example 9) using compound 24 (200 mg, 1.15 mmol), tert-butyl3-bromopropylcarbamate (273 mg, 1.15 mmol), Cs₂CO₃ (753 mg, 2.31 mmol)and DMF (5 mL). 235 mg (62% yield) of compound 25a were isolated. ¹HNMR(500 MHz, CDCl₃) δ PPM: 7.05-7.19 (m, 3H), 6.80 (dd, 1H), 4.7 (br, 1H),3.99 (t, 2H), 3.31 (t, 2H), 1.97 (m, 2H), 1.44 (s, 9H).

Synthesis of tert-butyl3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propylcarbamate

Compound 26a was prepared according to the procedure for compound 26(Example 9) using compound 25a (300 mg, 0.9 mmol),bis(Pinacolato)diboron (346 mg, 1.36 mmol), Palladium(II)acetate (12 mg,0.05 mmol), BINAP (22 mg, 0.03 mmol) and Cesium carbonate (590 mg,1.8mmol) in dioxane (20 mL. Crude compound 26a (350 mg) was used in thenext step without further purification.

Synthesis of tert-butyl3-(3-(9H-pyrido[2,3-b]indol-4-yl)phenoxy)propylcarbamate (27a)

Compound 27a was prepared according to the procedure for compound 27(Example 9) using compound 6 (100 mg, 0.40 mmol), compound 26a (229 mg,0.60 mmol), aqueous sodium carbonate (2 M, 1 mL, 2.0 mmol) and Pd(PPh₃)₄(23 mg, 0.02 mmol) in dioxane (4 mL). Compound 27a was isolated in 54%yield. Mass (m/z): 418.4 (M+H).

Synthesis of 3-(3-(9H-pyrido[2,3-b]indol-4-yl)phenoxy)propan-1-amine(28a, I-10)

Compound 28a was prepared according to the procedure for compound 28(Example 9) using compound 27a (100 mg, 0.23 mmol) and trifluoroaceticacid (136 mg, 1.2 mmol) in anhydrous dichloromethane (3 mL). The crudeproduct was purified by preparative HPLC giving compound 28a (13 mg, 17%yield) as an off white solid. ¹HNMR (500 MHz, DMSO-d₆) δ PPM: 11.97 (br,1H), 8.44(d, 1H), 7.80 (br, 2H), 7.51-7.54 (m, 2H), 7.39 (t, 1H), 7.25(d, 1H), 7.19 (s, 1H), 7.08 (d, 2H), 7.07 (dd, 2H), 7.02 (t, 1H); 4.12(t,2H), 2.99 (m,2H), 2.01 (t,2H). Mass (m/z): 318.3 (M+H). Purity:97.02% by HPLC.

Example 11 Scheme for the Preparation of Compounds 36a-d (I-11, I-13,I-14, I-12) and General Procedure for the Preparation of Compounds 32a-d

General Procedure for the Synthesis of Compounds 32a-d (I-11, I-13,I-14, I-12)

A mixture of 2,3-dichloro pyridine 29 (1 g, 6.76 mmol), an aniline 30(7.43 mmol), Pd(OAc)₂ (98 mg, 0.065 mmol), PPh₃ (198 mg, 0.13 mmol) andNaO^(t)Bu (780 mg, 1.2 mmol) in o-xylene (16 mL) was sparged withnitrogen for 5 min, placed under a nitrogen atmosphere, and heated to130° C. for 3 h in a sealed sample vial. The reaction mixture was cooledto room temperature giving compound 31. Pd(OAc)₂ (98 mg, 0.065 mmol),PCy₃ (200 mg, 0.1 mmol), DBU (2 g, 2 mmol) and dimethyl acetamide (16mL) were added to the reaction vessel. The reaction mixture was spargedfor 5 min, placed under a nitrogen atmosphere and heated to 160° C. for16 h. The reaction mixture was concentrated to dryness. The residue wasdissolved in ethyl acetate (2×200 mL). The mixture was washed with water(3×50 mL) and then brine (2×50 mL), dried over anhydrous Na₂SO₄,filtered and concentrated. The residue was purified on silica gel (70%EtOAc/hexane) giving compounds 32.

Example 12 Synthesis of 7-methyl-9H-pyrido[2,3-b]indole (32a, I-11)

Compound 32a was prepared according to Example 11 using 3-methylaniline(877 mg, 7.43 mmol) 30a and proportionate molar equivalents of compound29, Pd(OAc)₂, PPh₃, NaO^(t)Bu and o-xylene. Continuing according toExample 11, crude isolated compound 31a was converted to compound 32ausing proportionate molar equivalents of Pd(OAc)₂, PCy₃, DBU anddimethyl acetamide. 480 mg (40% yield) of compound 32a were isolated. ¹HNMR (500 MHz, DMSO-d₆) δ PPM: 11.61 (br, 1H), 8.41(dd, 2H), 8.02 (d,1H), 7.38 (s, 1H), 7.21 (d, 1H), 7.15 (d, 1H), 2.45 (s, 3H). Mass (m/z):183.2 (M+H).

Synthesis of 7-methyl-9H-pyrido[2,3-b]indole 1-oxide (33a)

Compound 33a was prepared according to the procedure for compound 5(Example 1) using compound 32a (370 mg, 2.02 mmol) and proportionatemolar equivalents of aqueous H₂O₂ (35%) and CH₃COOH. 160 mg (40% yield)of compound 33a were isolated. Mass (m/z): 199.2(M+H).

Synthesis of 4-bromo-7-methyl-9H-pyrido[2,3-b]indole (34a)

Compound 34a was prepared according to the procedure for compound 6(Example 1) using compound 33a (155 mg, 0.782 mmol) and proportionatemolar equivalents of anhydrous DMF and POBr₃. 61 mg (30% yield) ofcompound 34a were isolated. Mass (m/z): 261.1, 263.1.

Synthesis of tert-butyl 3-(7-methyl-9H-pyrido[2,3-b]indol-4-yl)phenethylcarbamate (35a)

Compound 35a was prepared according to the procedure for compound 8(Example 1) using compound 34a (61 mg, 0.23 mmol) and proportionatemolar equivalents of boronate ester 7, aqueous sodium carbonate (2.0M),dioxane and Pd(PPh₃)₄. 49 mg (50% yield) of compound 35a were isolated.Mass (m/z): 417.3(M+H).

Synthesis of 2-(3-(7-methyl-9H-pyrido[2,3-b]indol-4-yl)phenyl)ethanamine(36a, I-11)

Compound 36a was prepared according to the procedure for compound 9(Example 1) from compound 35a (49 mg, 0.117 mmol) and treating withproportionate molar equivalents of trifluoroacetic acid in anhydrousdichloromethane. Purification by preparative HPLC gave 14 mg (37% yield)of compound 36a. ¹H NMR (500 MHz, DMSO-d₆) δ PPM: 11.89 (br, 1H),8.39(d, 1H), 7.58 (d, 1H), 7.35 (m, 2H),7.05 (d, 1H), 6.86 (m, 3H), 6.80(d, 1H), 6.02(br, 1H), 2.98 (t, 2H),2.40 (s, 3H). ¹H NMR D₂O Exchange(500 MHz, DMSO-d₆) δ PPM: 8.32 (d, 1H), 7.50 (d, 1H), 7.35 (m, 2H), 7.05(d, 1H), 6.86 (m, 3H), 6.80 (d, 1H), 3.35 (t, 2H), 2.98 (t, 2H), 2.38(s, 3H). Mass (m/z): 317.3 (M+H).

Example 13 Synthesis of 6-methyl-9H-pyrido[2,3-b]indole (32b, I-13)

Compound 32b was prepared according to Example 11 using 4-methylaniline(877 mg, 7.43 mmol) 30b and proportionate molar equivalents of compound29, Pd(OAc)₂, PPh₃, NaO^(t)Bu and o-xylene. Continuing according toExample 11, crude isolated compound 31b was converted to compound 32busing proportionate molar equivalents of Pd(OAc)₂, PCy₃, DBU anddimethyl acetamide. 495 mg (41% yield) of compound 32b were isolated. ¹HNMR (500 MHz, DMSO-d₆) δ ppm: 11.59 (br, 1H), 8.41(d, 1H), 8.38 (d, 1H),7.9 (s, 1H), 7.38 (d, 1H), 7.25 (d, 1H), 7.18 (t, 1H), 2.42(s, 3H). Mass(m/z): 183.2 (M+H).

Synthesis of 6-methyl-9H-pyrido[2,3-b]indole 1-oxide (33b)

Compound 33b was prepared according to the procedure for compound 5(Example 1) using compound 32b (420 mg, 2.29 mmol) and proportionatemolar equivalents of aqueous H₂O₂ (35%) and CH₃COOH. 173 mg (38% yield)of compound 33b were isolated. Mass (m/z): 199.2 (M+H).

Synthesis of 4-bromo-6-methyl-9H-pyrido[2,3-b]indole (34b)

Compound 34b was prepared according to the procedure for compound 6(Example 1) using compound 33b (170 mg, 0.857 mmol) and proportionatemolar equivalents of anhydrous DMF and POBr₃. 69 mg (31% yield) ofcompound 34b were isolated. Mass (m/z): 261.1, 263.1.

Synthesis of tert-butyl3-(6-methyl-9H-pyrido[2,3-b]indol-4-yl)phenethylcarbamate (35b)

Compound 35b was prepared according to the procedure for compound 8(Example 1) using compound 34b (68 mg, 0.256 mmol) and proportionatemolar equivalents of boronate ester 7, aqueous sodium carbonate (2.0M),dioxane and Pd(PPh₃)₄. 54 mg (49% yield) of compound 35b were isolated.Mass (m/z): 417.3(M+H).

Synthesis ofN1-(3-(6-methyl-9H-pyrido[2,3-b]indol-4-yl)phenyl)ethane-1,2-diamine(36b, I-13)

Compound 36b was prepared according to the procedure for compound 9(Example 1) using compound 35b (53 mg, 0.126 mmol) and proportionatemolar equivalents of trifluoroacetic acid and anhydrous dichloromethane.Purification by preparative HPLC gave 16 mg (40% yield) of compound 36b.¹H NMR (500 MHz, DMSO-d₆) δ PPM: 11.95 (br, 1H), 8.62(d, 1H), 7.70(s,1H), 7.65 (d, 2H),7.58 (t, 1H), 7.45 (d, 1H), 7.24 (d, 1H), 7.10(dd,2H), 7.0 (d, 1H), 6.20 (br, 1H),3.20 (t, 2H), 2.70 (s, 3H). ¹H NMR D₂OExchange (500 MHz, DMSO-d₆) δ PPM: 8.38 (d, 1H), 7.45 (m, 2H), 7.38 (t,1H),7.22 (d, 1H), 7.05 (d, 1H), 6.85 (m, 2H), 6.80 (d 1H), 3.35(t, 2H),2.98 (t, 2H),2.22 (s, 3H). Mass (m/z): 317.3 (M+H).

Example 14 Synthesis of 6 chloro-9H-pyrido[2,3-b]indole (32c)

Compound 32c was prepared according to Example 11 using 4-chloroaniline(1.03 g, 7.43 mmol) 30c and proportionate molar equivalents of compound29, Pd(OAc)₂, PPh₃, NaO^(t)Bu and o-xylene. Continuing according toExample 11, crude isolated compound 31c was converted to compound 32cusing proportionate molar equivalents of Pd(OAc)₂, PCy₃, DBU anddimethyl acetamide. 400 mg (30% yield) of compound 32c were isolated. ¹HNMR (500 MHz, DMSO-d₆) δ PPM: 11.95 (br, 1H), 8.59(d, 1H), 8.42 (d, 1H),8.26 (s, 1H), 7.58(d, 1H), 7.49 (d, 1H), 7.21 (t, 1H). Mass (m/z):203.2, 205.2.

Synthesis of 6-chloro-9H-pyrido[2,3-b]indole 1-oxide (33c)

Compound 33c was prepared according to the procedure for compound 5(Example 1) using compound 32c (380 mg, 1.88 mmol) and proportionatemolar equivalents of aqueous H₂O₂ (35%) and CH₃COOH. 205 mg (50% yield)of compound 33c were isolated. Mass (m/z): 219.2, 221.2.

Synthesis of 4-bromo-6-chloro-9H-pyrido[2,3-b]indole (34c)

Compound 34c was prepared according to the procedure for compound 6(Example 1) using compound 33c (200 mg, 0.917 mmol) and proportionatemolar equivalents of anhydrous DMF and POBr₃. 90 mg (35% yield) ofcompound 34c were isolated. Mass (m/z): 283.0, 285.0.

Synthesis of tert-butyl 3-(6-chloro-9H-pyrido[2,3-b]indol-4-yl)phenethylcarbamate (35c)

Compound 35c was prepared according to the procedure for compound 8(Example 1) using compound 34c (85 mg, 0.302 mmol) and proportionatemolar equivalents of boronate ester 7, aqueous sodium carbonate (2.0M),dioxane and Pd(PPh₃)₄. 59 mg (45% yield) of compound 35c were isolated.Mass (m/z): 437.3, 439.3.

Synthesis ofN1-(3-(6-chloro-9H-pyrido[2,3-b]indol-4-yl)phenyl)ethane-1,2-diamine(36c, I-14)

Compound 36c was prepared according to the procedure for compound 9(Example 1) using compound 35c (58 mg, 0.133 mmol) and proportionatemolar equivalents of trifluoroacetic acid and anhydrous dichloromethane.Purification by preparative HPLC gave 15 mg (36% yield) of compound 36c.¹H NMR (500 MHz, DMSO-d₆) δ PPM: 12.10 (br, 1H), 8.40(d, 1H), 7.80(br,2H), 7.55 (s, 1H),7.50 (d, 1H), 7.40 (d, 1H), 7.38 (t, 1H), 7.08(d, 1H),6.8 (m, 3H), 6.00 (br, 1H), 3.00 (t, 2H). ¹H NMR D₂O Exchange (500 MHz,DMSO-d₆) δ PPM: 8.38 (d, 1H), 7.59 (m, 2H), 7.39 (d, 1H), 7.35 (t, 1H),7.15 (d, 1H), 6.89 (m, 3H), 3.38 (t, 2H), 2.98 (t, 2H). Mass (m/z):337.2. 339.2.

Example 15 Synthesis of 7-methoxy-9H-pyrido[2,3-b]indole (32d)

Compound 32d was prepared according to Example 11 using 3-methoxyaniline(890 mg, 7.43 mmol) 30d and proportionate molar equivalents of compound29, Pd(OAc)₂, PPh₃, NaO^(t)Bu and o-xylene. Continuing according toExample 11, crude isolated compound 31d was converted to compound 32dusing proportionate molar equivalents of Pd(OAc)₂, PCy₃, DBU anddimethyl acetamide. 355 mg (30% yield) of compound 32d were isolated. ¹HNMR (500 MHz, DMSO-d₆) δ PPM: 11.61 (br, 1H), 8.38(d, 1H), 8.34 (d,1H),7.18 (d, 1H), 6.90 (s, 1H), 6.81 (t, 1H), 3.82(s, 3H), Mass (m/z): 199.2(M+H).

Synthesis of 7-methoxy-9H-pyrido[2,3-b]indole 1-oxide (33d)

Compound 33d was prepared according to the procedure for compound 5(Example 1) using compound 32d (330 mg, 1.67 mmol) and proportionatemolar equivalents of aqueous H₂O₂ (35%) and CH₃COOH. 89 mg (25% yield)of compound 33d were isolated. Mass (m/z): 215.2 (M+H).

Synthesis of 4-bromo-7-methoxy-9H-pyrido[2,3-b]indole (34d)

Compound 34d was prepared according to the procedure for compound 6(Example 1) using compound 33d (88 mg, 0.411 mmol) and proportionatemolar equivalents of anhydrous DMF and POBr₃. 45 mg (40% yield) ofcompound 34d were isolated. Mass (m/z): 277.1, 279.1.

Synthesis of tert-butyl3-(6-methoxy-9H-pyrido[2,3-b]indol-4-yl)phenethyl carbamate (35d)

Compound 35d was prepared according to the procedure for compound 8(Example 1) using compound 34d (44 mg, 0.158 mmol) and proportionatemolar equivalents of boronate ester 7, aqueous sodium carbonate (2.0M),dioxane and Pd(PPh₃)₄. 27 mg (40% yield) of compound 35d were isolated.Mass (m/z): 433.3 (M+H).

SynthesisN1-(3-(7-methoxy-9H-pyrido[2,3-b]indol-4-yl)phenyl)ethane-1,2-diamine(36d, I-12)

Compound 36d was prepared according to the procedure for compound 9(Example 1) using compound 35d (27 mg, 0.062 mmol) and proportionatemolar equivalents of trifluoroacetic acid and anhydrous dichloromethane.Purification by preparative HPLC gave 6.3 mg (30% yield) of compound36d. Mass (m/z): 333.2 (M+H).

Additional compounds of formula I were prepared in a mannersubstantially similar to that described above.

Example 16 In Vitro CaMKIIδ Activity Assay

An exemplary procedure for the in vitro CaMKIIδ inhibition assay, whichcan be used to determine the inhibitory action of compounds of theinvention toward CaMKII, follows. The procedure is taken from Chao L H,et al., (2010) Nat Struct Mol Biol. 17(3): 264-272, which isincorporated herein by reference in its entirety.

The inhibition of CaMKII activity was evaluated using a coupled assaymeasuring ADP released following ATP hydrolysis and phosphor-transfer tothe peptide substrate AC3 (KKALHRQETVDAL; SEQ ID NO: 1) (1). A fulllength, C-terminal His/Gln tagged CaMKIIδ construct was used (sequencein Table 2 below).

TABLE 2 Amino acid sequence of CaMKIIδ construct.M A S T T T C T R F T D E Y Q L F E E L G K G A FS V V R R C M K I P T G Q E Y A A K I I N T K K LS A R D H Q K L E R E A R I C R L L K H P N I V RL H D S I S E E G F H Y L V F D L V T G G E L F ED I V A R E Y Y S E A D A S H C I Q Q I L E S V NH C H L N G I V H R D L K P E N L L L A S K S K GA A V K L A D F G L A I E V Q G D Q Q A W F G F AG T P G Y L S P E V L R K D P Y G K P V D M W A CG V I L Y I L L V G Y P P F W D E D Q H R L Y Q QI K A G A Y D F P S P E W D T V T P E A K D L I NK M L T I N P A K R I T A S E A L K H P W I C Q RS T V A S M M H R Q E T V D C L K K F N A R R K LK G A I L T T M L A T R N F S A A K S L L K K P DG V K E S T E S S N T T I E D E D V K A R K Q E II K V T E Q L I E A I N N G D F E A Y T K I C D PG L T A F E P E A L G N L V E G M D F H R F Y F EN A L S K S N K P I H T I I L N P H V H L V G D DA A C I A Y I R L T Q Y M D G S G M P K T M Q S EE T R V W H R R D G K W Q N V H F H R S G S P T VP I K L G S F L D H S F G A R A Q V X G H N H N H (SEQ ID NO: 2)

Compounds were added in 5 uL volume to wells in UV transparent 96-wellplates (½ area well size). The final compound concentrations testedranged from 0.5 nM to 10 uM). Assays were performed in duplicate.CaMKIIδ is added to at a final concentration of 16 nM to a mixturecontaining 100 mM Tris (pH 7.5), 150 mM KCl, 0.27 mM EGTA, 1.3 mM PEP,0.2 mg/ml AC3, 6.9% (v/v) PK/LDH mixture (Sigma P0294), 0.38 mM NADH andkept on ice. 72 uL of the enzyme mixture was added to the wellscontaining compounds and the plate was shaken briefly and kept on ice.The assay was initiated by adding 23 uL of a mixture containing 100 mMTris (pH 7.5), 150 mM KCl, 1.7 mM CaCl2, 48 mM MgCl2, 0.35 mM ATP and6.7 ug/mL calmodulin. The rate of ADP released was measured as the rateof absorbance decrease at 340 nM at 25° C. and plotted against the logof the compound concentration (FIG. 1). IC₅₀ data were fitted usingGraphPad Prism software.

The results of the in vitro CaMKIIδ activity assays are set forth inTable 3. The compound numbers correspond to the compound numbers inTable 1. Compounds having an activity designated as “A” provided anIC₅₀≦50 nM; compounds having an activity designated as “B” provided anIC₅₀ of 50-250 nM; compounds having an activity designated as “C”provided an IC₅₀ of 250-1000 nM; and compounds having an activitydesignated as “D” provided an IC₅₀≧1 μM. “NA” stands for “not assayed.”The enzyme inhibition curve for compound I-7 against CaMKIIδ is shown inFIG. 1.

TABLE 3 Results of in vitro CaMKII activity inhibition assays. CompoundID CaMKIIδ IC₅₀ I-1 B I-2 D I-3 D I-4 B I-5 C I-6 B I-7 A I-8 A I-9 AI-10 A I-11 A I-12 A I-13 A I-14 A

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments that utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments that have been represented by way of example.

We claim:
 1. A compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein: one of R^(2′)and R^(3′) is -L¹-R^(x), and the other is hydrogen; L¹ is a covalentbond or a straight or branched C₁₋₆ aliphatic group, wherein one or moremethylene groups are independently and optionally replaced by —NR^(a)—or —O—; R^(x) is selected from the group consisting of NH₂, guanidino,4-7 membered optionally substituted saturated heterocyclic ring having1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur,and 5-6 membered heteroaromatic ring having 1-2 heteroatomsindependently selected from sulfur, nitrogen and oxygen; each of R³, R⁴,R⁵, R⁶, R⁷, R⁸, R^(4′), R^(5′), and R^(6′) is independently selectedfrom the group consisting of hydrogen, halogen, —CN, —CF₃, —OR, —NR₂,—NO₂, —COOR, —CONR₂, and —R; each R^(a) is independently hydrogen orC₁₋₃ aliphatic; and each R is independently hydrogen or optionallysubstituted C₁₋₆ aliphatic.
 2. The compound of claim 1 of formula I-a:

or a pharmaceutically acceptable salt thereof, wherein: one of R^(2′)and R^(3′) is -L¹-R^(x), and the other is hydrogen; L¹ is a covalentbond or a straight or branched C₁₋₆ aliphatic group, wherein one or moremethylene groups are independently and optionally replaced by —NR^(a)—or —O—; R^(x) is selected from the group consisting of NH₂, guanidino,4-7 membered optionally substituted saturated heterocyclic ring having1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur,and 5-6 membered heteroaromatic ring having 1-2 heteroatomsindependently selected from sulfur, nitrogen and oxygen; each of R⁶ andR⁷ is independently selected from the group consisting of hydrogen,halogen, —CN, —CF₃, —OR, —NR₂, —NO₂, —COOR, —CONR₂, and —R; each R^(a)is independently hydrogen or C₁₋₃ aliphatic; and each R is independentlyhydrogen of optionally substituted C₁₋₆ aliphatic.
 3. The compound ofclaim 1 of formula II:

or a pharmaceutically acceptable salt thereof; wherein L¹ is a covalentbond or a straight or branched C₁₋₆ aliphatic group, wherein one or moremethylene groups are independently and optionally replaced by —NR^(a)—or —O—; R^(x) is selected from the group consisting of NH₂, guanidino,4-7 membered optionally substituted saturated heterocyclic ring having1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur,and 5-6 membered heteroaromatic ring having 1-2 heteroatomsindependently selected from sulfur, nitrogen and oxygen; each of R⁶ andR⁷ is independently selected from the group consisting of hydrogen,halogen, —CN, —CF₃, —OR, —NR₂, —NO₂, —COOR, —CONR₂, and —R; each R^(a)is independently hydrogen or C₁₋₃ aliphatic; and each R is independentlyhydrogen of optionally substituted C₁₋₆ aliphatic.
 4. The compound ofclaim 1 of formula III:

or a pharmaceutically acceptable salt thereof; wherein L¹ is a covalentbond or a straight or branched C₁₋₆ aliphatic group, wherein one or moremethylene groups are independently and optionally replaced by —NR^(a)—or —O—; R^(x) is selected from the group consisting of NH₂, guanidino,4-7 membered optionally substituted saturated heterocyclic ring having1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur,and 5-6 membered heteroaromatic ring having 1-2 heteroatomsindependently selected from sulfur, nitrogen and oxygen; each of R⁶ andR⁷ is independently selected from the group consisting of hydrogen,halogen, —CN, —CF₃, —OR, —NR₂, —NO₂, —COOR, —CONR₂, and —R; each R^(a)is independently hydrogen or C₁₋₃ aliphatic; and each R is independentlyhydrogen of optionally substituted C₁₋₆ aliphatic.
 5. The compound ofclaim 3 of formula IV:

or a pharmaceutically acceptable salt thereof; wherein R^(x) is selectedfrom the group consisting of NH₂, guanidino, 4-7 membered optionallysubstituted saturated heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen or sulfur, and 5-6 memberedheteroaromatic ring having 1-2 heteroatoms independently selected fromsulfur, nitrogen and oxygen; each of R⁶ and R⁷ is independently selectedfrom the group consisting of hydrogen, halogen, —CN, —CF₃, —OR, —NR₂,—NO₂, —COOR, —CONR₂, and —R; each R^(a) is independently hydrogen orC₁₋₃ aliphatic; and each R is independently hydrogen of optionallysubstituted C₁₋₆ aliphatic.
 6. The compound of claim 4 of formula V:

or a pharmaceutically acceptable salt thereof; wherein R^(x) is selectedfrom the group consisting of NH₂, guanidino, 4-7 membered optionallysubstituted saturated heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen or sulfur, and 5-6 memberedheteroaromatic ring having 1-2 heteroatoms independently selected fromsulfur, nitrogen and oxygen; each of R⁶ and R⁷ is independently selectedfrom the group consisting of hydrogen, halogen, —CN, —CF₃, —OR, —NR₂,—NO₂, —COOR, —CONR₂, and —R; each R^(a) is independently hydrogen orC₁₋₃ aliphatic; and each R is independently hydrogen of optionallysubstituted C₁₋₆ aliphatic.
 7. The compound of claim 3 of formula VI:

or a pharmaceutically acceptable salt thereof; wherein R^(x) is selectedfrom the group consisting of NH₂, guanidino, 4-7 membered optionallysubstituted saturated heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen or sulfur, and 5-6 memberedheteroaromatic ring having 1-2 heteroatoms independently selected fromsulfur, nitrogen and oxygen; each of R⁶ and R⁷ is independently selectedfrom the group consisting of hydrogen, halogen, —CN, —CF₃, —OR, —NR₂,—NO₂, —COOR, —CONR₂, and —R; and each R is independently hydrogen ofoptionally substituted C₁₋₆ aliphatic.
 8. The compound of claim 4 offormula VII:

or a pharmaceutically acceptable salt thereof; wherein R^(x) is selectedfrom the group consisting of NH₂, guanidino, 4-7 membered optionallysubstituted saturated heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen or sulfur, and 5-6 memberedheteroaromatic ring having 1-2 heteroatoms independently selected fromsulfur, nitrogen and oxygen; each of R⁶ and R⁷ is independently selectedfrom the group consisting of hydrogen, halogen, —CN, —CF₃, —OR, —NR₂,—NO₂, —COOR, —CONR₂, and —R; and each R is independently hydrogen ofoptionally substituted C₁₋₆ aliphatic.
 9. The compound of claim 3 offormula VIII:

or a pharmaceutically acceptable salt thereof; wherein R^(x) is selectedfrom the group consisting of NH₂, guanidino, 4-7 membered optionallysubstituted saturated heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen or sulfur, and 5-6 memberedheteroaromatic ring having 1-2 heteroatoms independently selected fromsulfur, nitrogen and oxygen; each of R⁶ and R⁷ is independently selectedfrom the group consisting of hydrogen, halogen, —CN, —CF₃, —OR, —NR₂,—NO₂, —COOR, —CONR₂, and —R; and each R is independently hydrogen ofoptionally substituted C₁₋₆ aliphatic.
 10. The compound of claim 4 offormula IX:

or a pharmaceutically acceptable salt thereof; wherein R^(x) is selectedfrom the group consisting of NH₂, guanidino, 4-7 membered optionallysubstituted saturated heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen or sulfur, and 5-6 memberedheteroaromatic ring having 1-2 heteroatoms independently selected fromsulfur, nitrogen and oxygen; each of R⁶ and R⁷ is independently selectedfrom the group consisting of hydrogen, halogen, —CN, —CF₃, —OR, —NR₂,—NO₂, —COOR, —CONR₂, and —R; and each R is independently hydrogen ofoptionally substituted C₁₋₆ aliphatic.
 11. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein the compound isselected from the following structures:


12. A pharmaceutical composition comprising a compound according toclaim 1 or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier, adjuvant, or vehicle.
 13. A methodof synthesizing a compound of the formula I-a:

wherein one of R^(2′) and R^(3′) is -L¹-R^(x), and the other ishydrogen; L¹ is a covalent bond or a straight or branched C₁₋₆ aliphaticgroup, wherein one or more methylene groups are independently andoptionally replaced by —NR^(a)— or —O—; R^(x) is selected from the groupconsisting of NH₂, guanidino, 4-7 membered optionally substitutedsaturated heterocyclic ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen or sulfur, and 5-6 memberedheteroaromatic ring having 1-2 heteroatoms independently selected fromsulfur, nitrogen and oxygen; each of R⁶ and R⁷ is independently selectedfrom the group consisting of hydrogen, halogen, —CN, —CF₃, —OR, —NR₂,—NO₂, —COOR, —CONR₂, and —R; each R^(a) is independently hydrogen orC₁₋₃ aliphatic; and each R is independently hydrogen of optionallysubstituted C₁₋₆ aliphatic; comprising: 1) contacting a compound of theformula:

wherein R⁶ and R⁷ are defined as above; with a compound of formula:

wherein R^(2′)-P and R^(3′)-P are as defined for R^(2′) and R^(3′) abovewith a primary amine protecting group on any primary amine; and 3)removing protecting group P.
 14. A method of treating a cardiovasculardisease, disorder, or condition, an inflammatory disease, disorder orcondition, a neurological disease, disorder or condition, an oculardisease, disorder or condition, a metabolic disease, disorder orcondition, a cancer or other proliferative disease, disorder orcondition, a bone disease, disorder or condition, or an addictivedisease, disorder, or condition, comprising administering to a patientin need thereof the composition according to claim
 12. 15. The method ofclaim 14 wherein the cardiovascular disease, disorder, or condition isselected from atrial fibrillation, ventricular arrhythmia, heartfailure, cardiac hypertrophy, atherosclerosis, restenosis; orcardiotoxicity arising from drug therapy, heart attack,ischemia-reperfusion injury, or catecholaminergic polymorphicventricular tachycardia.
 16. The method of claim 14 wherein theinflammatory disease, disorder or condition is asthma or rheumatoidarthritis.
 17. The method of claim 14 wherein the neurological disease,disorder or condition is pain or stroke.
 18. The method of claim 14wherein the metabolic disease, disorder, or condition is diabetes. 19.The method of claim 14 wherein the cancer or other proliferativedisease, disorder or condition is an osteosarcoma, a melanoma, or aprostate cancer.
 20. The method of claim 14 wherein the addictivedisease, disorder, or condition is opioid tolerance or dependence.